Long term effect of intraarticular adipose-derived stromal vascular fraction and platelet-rich plasma in dogs with elbow joint disease

doi:10.21203/rs.3.rs-3776760/v1

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Long term effect of intraarticular adipose-derived stromal vascular fraction and platelet-rich plasma in dogs with elbow joint disease

https://doi.org/10.21203/rs.3.rs-3776760/v1

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Background

Autologous stromal vascular fraction (SVF) with platelet rich plasma (PRP) may be used to treat osteoarthritis (OA) in dogs, but there is a lack of evidence regarding long time follow up after treatment. The aim of the current study was to examine long-term effects of a single intra-articular injection of SVF and PRP in dogs with confirmed elbow OA using orthopaedic lameness scoring and kinetic and kinematic gait analysis. For comparison of normal long-term variation of gait over time, a group of healthy control dogs (CD) was also evaluated.

Study design: Prospective longitudinal clinical study of 19 client owned dogs with elbow OA (OAD) treated with SVF and PRP and 8 CD not receiving treatment. Orthopaedic examinations, goniometry and seven kinetic and kinematic variables were evaluated before and six- and 12-months following treatment with SVF and PRP in the OAD group and twice with six months interval in the CD group. Orthopaedic examination variables (lameness, pain, range of motion) were scored and conflated to one out of three grades (normal, mildly or moderate/severely affected).

Results:

The OAD had an increase in fore-hind peak force symmetry 12 months after treatment (p< 0.05), but no other objective variables changed over time. Orthopaedic scores improved at six month follow up (p<0.05). None of the investigated gait variables had changed at six month follow up in the CD group.

Conclusions:

The current study did not show a clear benefit from SVF and PRP treatment in dogs with elbow OA.

Dog

Elbow dysplasia

PRP

Stemcell

Elbow joint dysplasia (ED), or abnormal joint development, is the most common cause of elbow osteoarthritis (OA) in dogs. Medial coronoid disease and humeral osteochondrosis are two significant causes of ED in dogs [1, 2]. Treatment of ED and secondary OA should optimally heal the defect in the cartilage and reverse the ongoing inflammatory process. Many medical and surgical therapies have been suggested, but with unclear long-term prognosis [3, 4]. Lameness and treatment effectiveness are frequently assessed using orthopaedic lameness scoring, kinetic gait analysis and owner questionnaires [5–7].

Mesenchymal stem cell therapy is an alternative or complementary treatment regime to NSAID, surgery and physiotherapy [8]. Adipose tissue in dogs contains multipotent stem cells of mesenchymal origin [9] and may be used to retrieve mesenchymal stem cells [10, 11]. The use of a stromal vascular fraction (SVF) allows harvest with minimal manipulation, reducing the requirement of in vitro expansion compared to culture of mesenchymal stem cells [12]. Methods for harvesting these cells in a SVF are commercially available; allowing for a one-step surgical procedure [13, 14]. One clinical trial and some case series report a decrease in stifle pain and improved cartilage growth (verified by magnetic resonance imaging or arthroscopy) after the application of SVF in humans [15]. In dogs, several clinical studies report positive short-term outcomes two to six months after autogenic SVF treatment. However, the methods used for evaluation vary amongst investigators; some used force plates for objective measurement and included double-blinded evaluations, while others had a subjective lameness score as the single evaluation method [16–20].

In dogs, mesenchymal stem cells with capability to develop into cartilage tissue have been harvested from adipose tissue [21]. Instead of repair with fibrocartilage, hyaline cartilage is formed in vivo after mesenchymal stem cell treatment, which may be important in joint disease [8]. The mechanism of action in vivo; from injection of stem cells to hyaline cartilage formation within the diseased joint, remains unclear. In humans, the mechanism has been suggested to be due to secretory effects from stem cells, direct differentiation of the stem cells in the joint, or a combination of both these effects [22].

There are significant amounts of growth factors and cytokines in platelet rich plasma (PRP), and it is the most accessible and affordable source of autologous cytokines and growth factors [23]. The use of PRP has been reported to inhibit the deleterious effects of inflammation and to counteract the development of OA with a chondroprotective effect in vivo in humans [24, 25]. Platelet rich plasma has been reported to improve lameness scores, reduce pain, and increase peak vertical forces in dogs with OA at three months follow up [26]. When PRP is activated, it may function as a scaffold for a SVF [27]. Based on owner questionnaires, improvements in dogs with hip dysplasia were reported in a double-blinded, placebo-controlled trial 24 weeks after receiving SVF and PRP treatment [14]. Currently, there is a lack of evidence regarding the long-term effects of SVF and PRP treatment in dogs with elbow OA.

The aim of the study was to evaluate the long-term effects of a single intra-articular injection of autologous SVF and PRP in dogs with confirmed elbow osteoarthritis (OA) using orthopaedic lameness scoring and kinetic and kinematic gait analysis. For comparison of normal variation in gait over time, a group of healthy control dogs (CD) was also evaluated.

The study was performed at the University Animal Hospital at the University of Agricultural Sciences, Uppsala, Sweden.

Dogs

Inclusion criteria: Dogs with elbow OA (OAD) and healthy CD were included in the study. Included OAD had to have a confirmed diagnose of unilateral or bilateral elbow OA based on diagnostic imaging (radiography or CT) findings, evaluated by a board-certified radiologist (ECVDI) or a veterinarian under their supervised training. To be eligible for enrollment, the OAD were required to previously have undergone NSAID treatment and rehabilitation including physical therapy exercises, with insufficient results and persistent lameness. Before or at the time of SVF/PRP therapy, dogs with OA could have received arthroscopic treatment; however, this was not a requirement for participation.

Clinically healthy dogs without a history- or clinical signs of systemic diseases, lameness or joint disease were included in the CD group.

All dogs included in the current study were also included in a study by Kjörk-Granström et al [28].

Exclusion criteria for the OAD included signs of systemic disease based on clinical examination and haematology/biochemistry blood profiles, or ongoing lameness not related to OA, such as due to an infectious process or an acute trauma. Within both groups, dogs were excluded if they were receiving medical treatments including immunosuppressive medications that potentially could mask pain responses. Within the CD group, any pain medication including NSAID was an exclusion criterion.

Treatment with NSAID was prohibited for the OAD group from 48 hours prior to and until two weeks following the SVF and PRP injection but was permitted as needed thereafter. Treatment with NSAID was registered (name of substance, dosage, duration of treatment), and the decision to initiate or stop ongoing therapy was decided by the responsible veterinarian (AB, MK). Due to long time follow up; not being able to treat with NSAID, if considered necessary, was considered unethical.

Clinical experimentation was approved by the Uppsala Ethics Committee (C102/15). Prior to participation, all owners gave their written consent.

Clinical evaluation at inclusion:

At the day of inclusion of all dogs, a complete health check was performed including physical evaluation. For the OAD, haematology and biochemistry blood profiles were evaluated to assure health and exclude signs of systemic disease. Additionally, elbow radiographs were taken, unless diagnostic imaging (CT or radiographs) had been performed less than six months prior to enrolment.

Orthopaedic evaluations:

All dogs were independently examined by a board-certified orthopaedic surgeon (AB) and a physical therapist (KP) at inclusion; before any potential treatment strategies had been implemented (all dogs), after six months (all dogs) and at least 12 months (OAD only) after enrolment into the study. A scoring system for clinical orthopaedic evaluations was used as described by Kjörk Granström et al [28]. Lameness at a walk and trot (graded 0–5), joint pain, palpable passive range of motion (PROM), goniometry and muscle measurements [29] were included in the scoring. Finally, when all data had been collected, but before results from the objective gait analysis were available, both examiners discussed their individual scorings and agreed on a common orthopaedic consensus score, into one of three categories; 1) normal-, 2) mildly- or 3) moderately- to severely affected (Table 1).

Table 1

Demographic data, orthopaedic lameness scoring, kinetic gait analysis and kinematic range of motion in 19 dogs with osteoarthritis (OAD), before treatment, 6 months and at least 12 months after stromal vascular fraction and platelet rich plasma treatment and 8 non lame (CD) dogs at 2 occasions with 6 months interval
	OAD at inclusion before treatment (n = 19)	OAD 6 months (n = 19)	OAD > 12 months (n = 11)	CD at inclusion (n = 8)	CD 6 months (n = 8)
Age (year)	4.2 ± 2.9	-	-	5.8 ± 2.8	-
Sex (female/male)	10/9	10/9	7/4	4/4	4/4
Bodyweight (kg)	35.3 ± 9.1	35.7 ± 9.7	33 ± 9.0	27.0 ± 6.6*	27.6 ± 6.6
BCS: U/N/O • U:1–3 under weight • N:4–5 normal weight • O: ≥6 overweight	0/5/14	0/4/15	0/1/10	0/4/4	0/4/4
NSAID (Yes/No), NSAID% Descriptive only	12/7 (63%)	5/14 (26%)	3/8 (37%)	0/8	0/8
Orthopaedic evaluations
Lameness score trot Presented with number of dogs in each lameness grade 0–5: (0/1/2/3/4/5)	Dx2/3/5/2/5/0 Sin2/4/3/4/3/0	Dx8/3/3/3/0/0 Sin5/9/0/2/0/0	Dx2/6/1/2/0/0 Sin4/2/3/0/0/0	All scored 0	All scored 0
Orthopaedic consensus score (normal/mild/ moderate&severe)	Dx 3/8/8 Sin 4/9/6	Dx 10/4/5# Sin 11/6/2#	Dx 3/6/2 Sin 7/3/1	Dx 8/0/0 Sin 8/0/0	Dx 8/0/0 Sin 8/0/0
Measured passive ROM elbow (degrees)	Dx 115.6 ± 11.3^a Sin 120.2 ± 7.3^a	Dx 16 ± 10.6^a Sin 119 ± 7.8^a	Dx 115 ± 10.5^a Sin 119 ± 10.7^a	Dx 125.6 ± 2.9^a Sin 125.4 ± 3.7^a	Dx 124.9 ± 2.5^a Sin 126.0 ± 2.5^a
Measured muscle mass (cm)	Dx 28.7 ± 2.5^a Sin 29.5 ± 2.9^a	Dx 29.1 ± 2.5^a Sin 9.6 ± 2.6^a	Dx 28.7 ± 3.0^a Sin 29.1 ± 2.8^a	Dx 26.6 ± 2.1^a Sin 27.0 ± 1.98^a	Dx 27.6 ± 2.1^a Sin 28.3 ± 2.1^a
Kinetic and kinematic evaluations
Total PF Symmetry	-7.1 ± 4.92^a*	-5.65 ± 3.8^a	-3.99 ± 1.74^a	-1.63 ± 1.58^a*	-2.98 ± 1.13^a
Fore-Hind PF Symmetry	-4.38 ± 3.44^a*	-3.29 ± 3.48^ab	-1.48 ± 2.75^b	-1.04 ± 1.51^a*	-1.55 ± 2.06^a
Fore PF Symmetry	0.52 ± 5.36^a	0.20 ± 3.3^a	-0.28 ± 1.25^a	0.24 ± 0.46^a	0.07 ± 0.32^a
Total Imp Symmetry	-5.17 ± 3.72^a*	-4.27 ± 3.26^a	-2.55 ± 1.38^a	-1.62 ± 0.61^a*	-1.82 ± 1.12^a
Fore-Hind Imp Symmetry	-1.71 ± 2.67^a*	-0.43 ± 2.86^a	-0.19 ± 1.70^a	0.55 ± 0.92^a*	0.38 ± 1.42^a
Fore Imp Symmetry	0.55 ± 5.58^a	-1.04 ± 3.94^a	-0.76 ± 1.73^a	-0.05 ± 0.90^a	0.68 ± 0.86^a
ROM Dx at walk	57.8 ± 6.1 ^a	55.6 ± 8.5 ^a	56.9 ± 6.6 ^a	54.5 ± 6.3^a	53.1 ± 5.1^a
ROM Sin at walk	58.2 ± 1.7 ^a	56.6 ± 9.2 ^a	58.3 ± 2.1 ^a	57.5 ± 2.7 ^a	54.8 ± 7.0^a
Symmetry Squares blinded evaluation: worse/unchanged/improved	Baseline (n = 19)	5/5/9 (n = 19)	1/3/7 (n = 11)	Baseline (n = 8)	0/7/1
Only treated joints are included in the table. Values are reported as mean and standard deviation. Values with * are significantly different between the OAD group and the CD group at the first occasion Values with # are significantly different for OAD before treatment and at six months (p < 0.05).
Within each row of continuous data, for OAD and CD respectively, values with the same superscript letter did not differ significantly (p > 0.05). The unilateral healthy limb is not presented in the orthopaedic evaluations. NSAID = Non steroid anti-inflammatory drug, BCS = Body condition score, ROM = Range of motion, PF = Peak Force, Imp = Impulse, Sin = sinister, Dx = dexter

Kinetic and kinematic evaluations

Kinetic and kinematic measurements were performed at the same intervals as the orthopaedic evaluations. The data collection and processing used are identical to those described by Kjörk Granström et al (Additional file 1) [28].

Gait velocity was measured in m/s (Total Peak Force (PF) Symmetry (Sym), Fore-Hind PF Sym, Fore PF Sym, Total Impulse(I) Sym, Fore-Hind I Sym, Fore I Sym and Kinematic Range of Motion (ROM) elbow). All the kinetic variables were normalized for body weight. Speed was normalized to mean speed at each session for each dog. Based on PF and I, the distribution of forces between all four legs was presented for each dog in a symmetry graph “Symmetry Squares” (SS) (Additional file 2) [28]. Diagonal forces represent symmetries between contralateral fore- and hindlimbs. Blinded evaluations of SS-graphs were reviewed independently by three clinicians to evaluate any change of symmetry forces (lameness detection) and any change over time, at the same intervals as the orthopaedic evaluations for the CD and OA group. All occasions for one individual were presented in the same graph and was graded as unchanged, improved or deteriorated over time (Fig. 1).

PRP preparation

Before induction of anaesthesia, 16–18 ml of venous blood was collected under strict aseptic conditions. The PRP was prepared with a validated method following MediVets In-Clinic Procedure Manual (Footnote I).

Preparation of SVF:

General anaesthesia was induced with propofol (Propovet Multidose, Zoetis, Finland), and isoflurane (IsoFlu, Zoetis, Finland) was used for maintenance. Adipose tissue (approximately 20 g) from the thoracic area was collected surgically using standard surgical procedures in the three first dogs included in the study. Due to seroma formation in two of the three dogs, abdominal falciform fat was collected from the remaining dogs without complications thereafter. Samples were processed in-house, using a commercial kit to yield SVF pellets according to the manufacturer´s instruction (Medi Vet America Manual, Footnote a). The final cell suspension with the ASC pellet and 2 mL of PRP was placed into a photobiostimulation unit (Medivet Stem Cell LED Activator ML-1) for 20 minutes before injection into the elbow joint under strict aseptic conditions. The adipose-derived cell population is activated by irradiating the cells with the ML-1, which generates certain wavelengths of light in the visible light spectrum to stimulate growth and differentiation of cells (Additional file 3). In bilaterally affected dogs; both elbows were treated with SVP and PRP.

Cell counts of SVF and PRP

Total nucleated cell count per gram adipose tissue was estimated for all dogs by laser flow cytometry of the final SVF solution (Footnote II) by diluting 0.1 ml of cell suspension with 0.4 ml normal saline in an empty vacutainer blood collection tube and placing it in the analyser.

Statistical evaluations

Statistical analyses were performed using a commercially available software program.^III Group data are presented as mean and standard deviation (SD). Data was analyzed using descriptive- as well as inferential statistics. Potential differences in baseline continuous outcome biomechanical variables (listed in Table 1) between the OAD and CD groups, as well as potential differences in the clinical orthopaedic consensus score between the first and the six month follow up examinations (for both the OAD and the CD dogs) were investigated using the Wilcoxon rank sum test. For the continuous outcome biomechanical variables listed in Table 1, differences between time-points of examinations (before treatment, six and ≥ 12 months, respectively, after treatment for the OA group, and six months apart for the CD group) were investigated separately for the OAD and CD groups using a mixed (repeated) linear model including dog identity as a random variable and time-point of examination. The normality of the distributions of model residuals were ensured by visual inspection. The Tukey HSD test was used as post-hoc test for pairwise comparisons of variables found to be significant in the overall analysis. A value of p ≤ 0.05 was considered significant for the analyses.

Dogs

The OAD group consisted of 19 dogs and included the following breeds: Belgian Shepherd (Malinois) (n = 1), Bernese Mountain dog (n = 2), German Shepherd (n = 3), German Spaniel (n = 1), Labrador Retriever (n = 7), Leonberger (n = 1), mixed breeds (n = 2), Rottweiler (n = 1), and Bullmastiff (n = 1). The CD group consisted of eight dogs and included the following breeds: German shepherd (n = 1), German wirehaired pointer (n = 1), Labrador Retriever (n = 3), mixed breed (n = 1), Rottweiler (n = 1) and Smooth collie (n = 1). Bodyweight was significantly lower in the CD group (Table 1).

Seventeen OAD had radiographic findings of OA including osteophytes, two of them with visible ED lesions. In two other dogs, ED with OA were confirmed through CT scan. Five OAD had unilateral elbow OA and 14 had bilateral elbow OA disease. The unilateral elbow disease was left sided in two dogs and right sided in three dogs. See Table 1 for further information regarding demographic data of included dogs.

Fifteen dogs were arthroscopically treated in one or both elbows before stem cell therapy. Three dogs had their second elbow arthroscopy and stem cell therapy at the same anaesthetic occasion. The contralateral elbow had been arthroscopically treated earlier in these three dogs. In one OAD dog, no arthroscopy had been performed. Two dogs (10%) developed short term complications from the treated elbows after treatment, both resolving spontaneously within three to four days; one developed a local swelling in the treated elbow and one showed pain from the elbow.

Treatment response and cell counts of SVF

Table 1 presents NSAID treatment, orthopaedic lameness scoring, orthopaedic consensus score, kinetic gait analysis and kinematic range of motion for OAD and CD. Nineteen dogs were evaluated at six months and 11 (58%) at ≥ 12 months Mean time from treatment at ≥ 12 months follow up was 1.5 years ± 0.56. The dog owners’ explanation for why eight dogs did not participate at ≥ 12 months are presented in Table 2. The clinical orthopaedic consensus score for the right and left front limb had changed six months after treatment in the OAD groups (p < 0.05 for both the right and the left front limb) (Table 1). Due to loss of 8/19 OAD at ≥ 12 months follow-up, statistical comparison was not performed for the categorical data for this examination occasion. Before treatment, Total PF and Total I, and Fore Hind PF and -I symmetry forces were lower in the OAD group compared to the CD group; Fore-Hind PF Symmetry increased at ≥ 12 months follow up in the OAD group compared to before treatment (p = 0.007) (Table 1).

Table 2

Information regarding the eight OAD (elbow osteoarthritis) that was not included in the 12 months follow up evaluation
Dog identity	Included at 12 months Yes/No	Cause for not being included at 12 month follow up and information from owner phone calls
OA1	Yes	-
OA2	Yes	-
OA3	No	Owner unwilling to participate at 12 months due to long traveling distance. According to owner, front limb lameness was observed after playing with the new puppy at that time, likely caused by OA.
OA4	Yes	-
OA5	Yes	-
OA6	Yes	-
OA7	Yes	-
OA8	Yes	-
OA9	No	Owner unwilling to participate at 12 months due to long traveling distance. According to owner, examination by local veterinarian showed that the dog had pain from metacarpophalangeal joints at 12 months
OA10	No	Chronic kidney disease, had a long period with good function (less pain and lameness) between six and 12 months after treatment
OA11	No	Euthanized due to elbow OA pain
OA12	Yes	-
OA13	Yes	-
OA14	Yes	-
OA15	Yes	-
OA16	No	Owner unwilling to participate at 12 months due to long traveling distance
OA17	No	Shoulder pain developed after trauma at 10 months, excluded from further evaluations. According to the orthopaedic subjective evaluation elbows where not painful (examination by AB)
OA18	No	Lumbar pain developed after six but before 12 months, diagnosed at the university animal hospital. Excluded from further evaluations, did not improve in elbow pain after six months according to owner or clinicians.
OA19	No	Euthanized due to lymphoma
OA = osteoarthritis

Results from the blinded evaluation of any changes in symmetry forces (SS graphs) over time is presented in Table 1. The eleven remaining OAD did not change between categories (worse/unchanged or improved) between six and ≥ 12 months follow up, meaning that if they were improved at six months, they had maintained that improvement at ≥ 12 months. The CD at six months were assessed as unchanged in 7/8 dogs. One dog was considered to put more weight on the front limbs at six months. The SS of four individual dogs is presented as examples in Fig. 1, and the Fore-Hind PF Sym index over time in all dogs is graphically presented in Fig. 2.

Cell counts of nucleated cells in the canine SVF was 8.9x106 (± 3.13 x 106) cells/gram of adipose tissue.

The study showed improvements in clinical orthopaedic consensus score six months after treatment and in Fore-Hind PF symmetry in the remaining 11 OAD ≥ 12 months after treatment. Other variables did not change statistically at six or ≥ 12 months, which means that the study was not able to confirm if SVF/PRP treatment is efficient in dogs with elbow OA. Ten percent of the dogs developed minor side effects after treatment.

The SVF/PRP did not show a clear benefit after six or ≥ 12 months. Reasons may vary, such as no/limited effect of therapy, only short time effect of therapy or not detectable changes due to too few included dogs. Also, the technique of the SVF procedure may have had an influence on the results. The procedure with SVF/PRP injection following an arthroscopic procedure, for instance, may not be ideal because of the risk of dilution. This has not yet been investigated.

The current study reports a longer follow up time compared to several other stem cell studies, however in a small sample of dogs. Due to the loss of follow up of 8/19 OAD at ≥ 12 months, categorical data, such as orthopaedic consensus score, was only evaluated at six months. Of the eight missing dogs, only one was reported to be excluded due to elbow OA (OAD11). Information presented in Table 2 shows that several dogs however were likely to have clinical problems due to elbow OA. Vilar et al reported a short-term (three months) positive effect after treatment with cultured stem cells without PRP in dogs with hip OA on objective gait analysis before relapse to levels before treatment at 180 days after treatment [30]. The clinical use of SVF or cultured cells is sparsely studied, but has been reported in one study by Marx et al, were SVF was reported to show more clearly positive clinical results compared to cultured cells [19]. The possible reasons for this is unknown, but SVF also contains other cells, which may be important for the healing process in the OA joint.

In human research, PRP has been concluded to enhance stem cell therapy [31], and this may also apply for dogs. In a blinded study including 39 dogs, evaluated with subjective lameness scoring after treatment, Cuervo et al [18] also reported that PRP may increase the duration of clinical effect with SVF. In contrary to our study, they did not include objective gait analysis.

Scoring of lameness and gait analysis measurements in dogs with mild and/or bilateral lameness, will be of lower magnitude, making the possibility to evaluate changes over time more difficult. As can be seen in Fig. 2, some individuals from the OAD had similar Fore-Hind PF Sym as CD, which clinically could be identified as normal variation in gait. Vilar et al reported an increase in ground reaction forces (GRF) in the more severely lame limb and a decrease in GRF of the less lame limb after stem cell therapy [20]. The lameness scores as presented in Table 1 included eight elbows with grade 4/5 before treatment. The lameness score severity decreased over time, and at six and 12 months follow up, no dog showed 4/5 in lameness score. However, due to few cases as well as loss of eight dogs at 12 months follow up, the lameness grading was only reported descriptively. The SS evaluation also identified an increase in forces in one or two front limbs in 47% of the OAD at six months, 63% of the OAD at 12 months, and 12% (one dog) of the CD at six months. A bias not related to treatment could result in the decrease of lameness severity over time. Bilateral lameness is considered to be more difficult to evaluate. The SS graphs will be useful for the physician as a tool to graphically discern how forces are distributed and changed over time, also seen in the current investigation. (Fig. 1) It was not possible to detect a difference in outcome after treatment between uni- or bilaterally lame dogs. As elbow OA is considered a chronic disease; it may wax and wane but generally gets worse over time, with an increase of lameness and NSAID therapy. Before SVF and PRP treatment, all treated dogs periodically had been treated with NSAID and rehabilitation, but the effect was considered clinically insufficient according to owner and veterinarian. The usage of NSAID decreased from 67% before treatment to 26% at six months. After 12 months, only 11 dogs were included and at that time; 37% received NSAID. It is possible that SVF and PRP therapy may reduce the need for NSAID in dogs, but further research, preferably double-blinded studies is needed to verify this finding as the owners might be biased from the treatment.

Based on the gait analysis results presented in Fig. 2, some of the young, severely lame dogs treated with SVF/PRP (OAD 6, 7, 8, and 14) may have benefited more from the therapy compared to older and severely lame dogs (OAD 3, 9, and 10). Such a potential difference in treatment effect between young and old dogs should be further investigated. An influence of age on the activity and number of human stem cells have been discussed [25]. Furthermore, chronicity of the joint disease in older dogs may have had an influence on SVF and PRP treatment effect.

Limitations:

The main limitation of the study was the relative low number of individuals included, and the loss of 8/12 OAD at 12 months follow up which may affect the results. There was also a lack of a placebo treated group of dogs, and the clinical evaluations were not blinded to treatment group or control group. This may cause a risk for bias towards clinical improvement in the treated OAD. Also, all orthopaedic evaluations are subjective and prone to bias [32].

In the current study, the number of administered active stem cells is undefined. Fresh SVF are heterogeneous with different types of cells and may be difficult to evaluate accurately due to debris. However, Maki et al. reported no difference in effect when three different concentrations of adipose mesenchymal stem cells were administered [33].

The current study did not show a clear benefit from SVF/PRP treatment in dogs with elbow OA. The investigated gait variables did not change over time in the CD group. Future research is needed to evaluate if young dogs benefit better from SVF/PRP compared to older dogs.

Authors’ contributions

AB, MA, MG and KP contributed to the initial study design. AB, MG and KP collected the data. MA performed extraction of SVF and preparation of PRP. IL performed the data analysis and LR performed the data-processing and analysis of the objective measurements. AB wrote the manuscript with substantial input from all co-authors. All authors read and approved the final manuscript.

Acknowledgements

The authors would like to acknowledge the invaluable assistance of Christoffer Roepstorff with the practical assistance during data collection for the objective gait analysis. The authors also wish to acknowledge all dogs and owners participating in the study.

Competing interests

The authors declare that they have no competing interests.

Availability of data and materials

The datasets used and analyzed during the current study are available from the corresponding author on reasonable request.

Consent for publication

All participating owners signed consent of statement.

Ethics approval

The study was ethically approved by the Uppsala animal ethics committee (C102/15). All dog owners provided signed informed consent prior to enrolment in the study.

Prior publication

Data have not been published previously.

Funding

This study was funded by Agria Pet Insurance Research Foundation. The founding covered all costs related to the study such as reflective markers etc, but the founder had no influence on the manuscript.

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Long term effect of intraarticular adipose-derived stromal vascular fraction and platelet-rich plasma in dogs with elbow joint disease

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Abstract

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Background

Methods

Dogs

Clinical evaluation at inclusion:

Orthopaedic evaluations:

Kinetic and kinematic evaluations

PRP preparation

Preparation of SVF:

Cell counts of SVF and PRP

Statistical evaluations

Results

Dogs

Treatment response and cell counts of SVF

Discussion

Limitations:

Conclusions

Declarations

References

Endnotes

Supplementary Files

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