Characterization of platelet rich plasma in feline immunodeficiency virus-infected cats: Cell, and PDGF-BB and TGF-ß1 growth factor analysis.

Autologous platelet-rich plasma (PRP) contains growth factors (GFs) that modulate the expression of inflammatory cells; thus, these products could be considered a good strategy to favor tissue regeneration in feline immunodeficiency (FIV) positive cats. However, there is no scientific documentation on obtaining PRP in FIV-positive cats. Authors hypothesized that PRP can be obtained in FIV cats following the PRGF®-Endoret® methodology. The objectives of this study were to compare the platelet, erythrocyte, and leukocyte concentration between whole blood (WB) and the PRP; and determine the concentration of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor ß1 (TGF-ß1) in FIV-positive cats. Sixteen adults FIV-positive asymptomatic cats were included in the study. WB samples were drawn and the PRP was obtained by centrifugation at 265g for 10 min. Erythrocyte and leukocyte, platelets, and mean platelet volume (MPV) were determined both in WB and in PRP. PDGF-BB and TGF-ß1 concentrations were additionally determined in PRP. Platelet concentration increased 1.1 times in PRP fraction compared to WB, but no significant differences were reported. MPV was statistically higher in WB than in PRP (p = 0.001). Erythrocytes and leukocytes counts were decreased by 99% and 92%, respectively in the PRP fraction (p < 0.001). Regarding TGF-ß1, a higher concentration was shown in the PRP (p < 0.02). Although the product obtained could not be classified as PRP according to the PRGF®-Endoret® methodology, based on the drastic reduction of RBC and WBC, the PLT concentrate is of high purity.

Intra-osseous plasma rich in growth factors enhances cartilage and subchondral bone regeneration in rabbits with acute full thickness chondral defects: Histological assessment.

Background: Intra-articular (IA) combined with intra-osseous (IO) infiltration of plasma rich in growth factors (PRGF) have been proposed as an alternative approach to treat patients with severe osteoarthritis (OA) and subchondral bone damage. The aim of the study is to evaluate the efficacy of IO injections of PRGF to treat acute full depth chondral lesion in a rabbit model by using two histological validated scales (OARSI and ICRS II). Methodology: A total of 40 rabbits were included in the study. A full depth chondral defect was created in the medial femoral condyle and then animals were divided into 2 groups depending on the IO treatment injected on surgery day: control group (IA injection of PRGF and IO injection of saline) and treatment group (IA combined with IO injection of PRGF). Animals were euthanized 56 and 84 days after surgery and the condyles were processed for posterior histological evaluation. Results: Better scores were obtained in treatment group in both scoring systems at 56- and 84-days follow-up than in control group. Additionally, longer-term histological benefits have been obtained in the treatment group. Conclusions: The results suggests that IO infiltration of PRGF enhances cartilage and subchondral bone healing more than the IA-only PRGF infiltration and provides longer-lasting beneficial effects.

Evaluation of Platelet-Rich Plasma by means of PRGF®-Endoret® protocol in leukemia cats: PDGF-BB and TGF-ß1 valuation.

Introduction: Feline leukemia virus (FeLV) is a chronic disease that leads to the weakening of a cat's immune system. Platelet-rich plasma (PRP) offers therapeutic effects for multiple diseases, the use of PRP and growth factors (GFs) determination could be an alternative treatment to improve the quality of life in these patients. The objectives of this study were to determine and compare the concentration of platelets (PLTs), red blood cells (RBCs) and white blood cells (WBCs) between samples of whole blood (WB), PRP and platelet-poor plasma (PPP) fractions, and to evaluate the concentration of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor ß1 (TGF-ß1) in both fractions in FeLV cats using a PRGF®-Endoret® protocol previously standardized in this species. Methods: WB was collected from 11 asymptomatic FeLV-positive cats. PRP and PPP was obtained following PRGF®-Endoret® technology according to centrifugation at 265 g for 10 min. Cellular components, RBCs, WBCs, PLTs, and the PDGF-BB and TGF-ß1 concentrations in PRP and PPP fractions were determined. Results: PLT in the PRP fraction was statistically higher than WB and PPP fraction, with no statistical differences between WB and PPP. PLT concentration increased 1.4 times in PRP fraction compared to WB. Mean platelet volume (MPV) did not differ significantly between the WB, PRP, and PPP fractions. Compared to WB, the absolute numbers of RBCs and WBCs were decreased by 99% and more than 95% in the PRP and PPP fractions, respectively. TGF-ß1 concentrations increased in PRP vs. PPP, with no changes in PDGF-BB. Discussion: Based on the degree of PLT enrichment and the absence of RBCs and WBCs, this blood product could be classified as a Pure Platelet-Rich Plasma (P-PRP). The presence of GFs in PRP and PPP samples suggests that the PRGF®-Endoret® methodology is suitable for obtaining PRP in FeLV cats, despite future studies are necessary to optimize the technique, standardize the results and assess clinical efficacy.

Corrigendum: Evaluation of a Standardized Protocol for Plasma Rich in Growth Factors Obtention in Cats: A Prospective Study.

[This corrects the article DOI: 10.3389/fvets.2022.866547.].

Evaluation of a Standardized Protocol for Plasma Rich in Growth Factors Obtention in Cats: A Prospective Study.

Introduction: Platelet-rich plasma (PRP) is an autologous plasma with platelet (PLT) concentration above that of whole blood (WB). PLTs contain growth factors (GFs) that promote tissular repair. Objectives: To determine and compare the concentrations of PLT, red blood cells (RBC) and white blood cells (WBC) between WB samples, PRP and platelet poor plasma (PPP) samples; and to analyze the concentrations of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor ß1 (TGF-ß1) in the PRP and PPP of healthy adult cats using a standardized protocol with PRGF®-Endoret® characteristics. Material and Methods: WB was collected from 30 cats. PRP was obtained following three centrifugation protocols using PRGF®-Endoret® technology: 255, 260, and 265 g for 10 min each. The cellular components, RBC, WBC, PLT, and the concentrations of PDGF-BB and TGF-ß1 in the PRP and PPP fractions were determined for each protocol. Results: PLTs in the PRP fraction were statistically higher than WB, with no statistical differences between PPP and WB. In PRP fraction, PLT concentration was increased 1.4 times on average at 255 g; 1.3 times at 260 g and, 1.5 times at 265 g without statistical differences among them. The mean platelet volume (MPV) was significantly higher in WB compared to PRP and PPP fractions without significant differences between protocols. Compared to WB, the number of RBCs and WBCs was reduced by 99% and by more than 95% in PRP and PPP respectively, without significant differences between protocols. PDGF-BB concentrations were statistically higher in PRP than in PPP fractions, however, TGF-ß1 concentrations did not vary between fractions at 260 g. Comparing the three protocols within PRP and PPP fractions, no differences in PDGF-BB and TGF-ß1 concentrations were observed. Clinical Relevance: The study shows scientific evidence regarding the obtention of PRP in cats using the PRGF®-Endoret® technology for the quantification of PDGF-BB and TGF-ß1. At 265 g for 10 min, PLT concentration was increased 1.5 times with unnoticeable erythrocytes and leukocytes in the samples. These results clearly show that the PRGF®-Endoret® methodology is suitable to obtain PRP in cats. Further studies are needed to determine the clinical efficacy of the obtained PGRF in the treatment of different pathologies in cats.

Intra-osseous infiltration of adipose mesenchymal stromal cells and plasma rich in growth factors to treat acute full depth cartilage defects in a rabbit model: Serum osteoarthritis biomarkers and macroscopical assessment.

Introduction: Intra-articular infiltration of plasma rich in growth factors (PRGF) and adipose mesenchymal stromal cells (AMSCs) are known to inhibit osteoarthritis progression. However, in severely affected patients, the treatment cannot reach the deeper layers of the articular cartilage; thus, its potential is limited. To overcome this limitation, intra-osseous infiltrations have been suggested. The purpose of this study is to assess the impact of intra-osseous infiltration therapies on serum biomarkers of osteoarthritis and to assess cartilage regeneration macroscopically. Materials and methods: A total of 80 rabbits were divided into four groups based on the intra-osseous treatment administered on the day of surgery: control, PRGF, AMSCs and a combination of PRGF + AMSCs. In addition, all groups received a single intra-articular administration of PRGF on the same day. Serum biomarker levels were measured before infiltration and 28-, 56-, and 84-days post infiltration, and macroscopical assessment was conducted at 56- and 84-days follow-up post infiltration. Results: In the PRGF + AMSCs group, significantly lower concentrations of hyaluronic acid and type II collagen cleavage neoepitope were recorded at all time points during the study, followed by PRGF, AMSCs and control groups. Regarding macroscopical assessment, lower scores were obtained in PRGF + AMSCs group at all study times. Discussion: The results suggest that the combination of intra-articular PRGF with intra-osseous PRGF or AMSCs achieves better results in rabbits with acute chondral defects and that intra-osseous infiltration is a safe procedure.

Treating Full Depth Cartilage Defects with Intraosseous Infiltration of Plasma Rich in Growth Factors: An Experimental Study in Rabbits.

OBJECTIVE: Intraarticular (IA) administration of platelet-rich plasma (PRP) has been proposed as a new strategy to halt osteoarthritis (OA) progression. In patients with severe OA, its potential is limited because it is unable to reach the subchondral bone, so a new strategy is needed, and intraosseous (IO) infiltration has been suggested. The purpose is to assess the impact of IA together with IO infiltration of plasma rich in growth factors (PRGF) in serum hyaluronic acid (HA) and type II collagen cleavage neoepitope (C2C) levels. DESIGN: A total of 32 rabbits were included in the study and randomly divided into 2 groups: control and treatment. A 4-mm chondral defect was created in the medial femoral condyle and IA followed by IO infiltration were performed. Serum C2C and HA levels were measured using enzyme-linked immunosorbent assay (ELISA) tests before infiltration and 28, 56, and 84 days post-infiltration. RESULTS: Significant lower C2C serum levels were obtained in treatment group (IA + IO infiltration of PRGF) at 84 days post-infiltration than in control group (IA infiltration of PRGF + IO infiltration of saline solution), while no significant differences between groups were reported at any other study times. Regarding HA, at 56 days post-infiltration, greater significant levels were seen in the treatment group. However, at 84 days post-infiltration, no significant differences were obtained, although lower levels were reported in the treatment group. CONCLUSIONS: Despite inconclusive, the results suggest that the combination of IA and IO infiltration with PRGF may enhance cartilage and subchondral bone regeneration, but further studies are needed.

Cell and Cell Free Therapies in Osteoarthritis.

Osteoarthritis (OA) is the most common articular disease in adults and has a current prevalence of 12% in the population over 65 years old. This chronic disease causes damage to articular cartilage and synovial joints, causing pain and leading to a negative impact on patients' function, decreasing quality of life. There are many limitations regarding OA conventional therapies-pharmacological therapy can cause gastrointestinal, renal, and cardiac adverse effects, and some of them could even be a threat to life. On the other hand, surgical options, such as microfracture, have been used for the last 20 years, but hyaline cartilage has a limited regeneration capacity. In recent years, the interest in new therapies, such as cell-based and cell-free therapies, has been considerably increasing. The purpose of this review is to describe and compare bioregenerative therapies' efficacy for OA, with particular emphasis on the use of mesenchymal stem cells (MSCs) and platelet-rich plasma (PRP). In OA, these therapies might be an alternative and less invasive treatment than surgery, and a more effective option than conventional therapies.

Pedobarography: a novel approach to test the efficacy of treatments for lameness; an experience with mavacoxib in dogs with elbow osteoarthritis.

BACKGROUND: Pedobarographic analyses detect pressure redistribution among limbs and within limbs in humans, equids and dogs. The main objective of this study was to assess the usefulness of a set of pedobarographic parameters for the detection of lameness, as well as for its suitability for assessing the effects of therapies against osteoarthritis in dogs. With this purpose, eleven large-breed lame dogs with unilateral osteoarthritis due to elbow dysplasia were evaluated using a pressure platform prior to (D0) and after 3 months (D90) of treatment with mavacoxib, a COX-2 selective NSAID. The obtained parameters were: pressure distribution between lame and sound limbs, as well as paw area, mean pressure, and peak pressure of both lame and sound limbs. RESULTS: The results showed statistical differences in all these parameters between lame and sound limbs at D0; however, at D90, differences were significantly decreased as result of the treatment, indicating a substantial functional recovery under the study design conditions. CONCLUSIONS: The provided data prove the suitability of this novel technique in canine models for the quantitative and objective assessment of lameness, but also for the evaluation of treatments for lameness caused by articular pain.

Adipose-Derived Mesenchymal Stem Cells: A Promising Tool in the Treatment of Musculoskeletal Diseases.

Chronic musculoskeletal (MSK) pain is one of the most common medical complaints worldwide and musculoskeletal injuries have an enormous social and economical impact. Current pharmacological and surgical treatments aim to relief pain and restore function; however, unsatiscactory outcomes are commonly reported. In order to find an accurate treatment to such pathologies, over the last years, there has been a significantly increasing interest in cellular therapies, such as adipose-derived mesenchymal stem cells (AMSCs). These cells represent a relatively new strategy in regenerative medicine, with many potential applications, especially regarding MSK disorders, and preclinical and clinical studies have demonstrated their efficacy in muscle, tendon, bone and cartilage regeneration. Nevertheless, several worries about their safety and side effects at long-term remain unsolved. This article aims to review the current state of AMSCs therapy in the treatment of several MSK diseases and their clinical applications in veterinary and human medicine.

Center of pressure limb path differences for the detection of lameness in dogs: a preliminary study.

BACKGROUND: The limb center of pressure (COP) path measures and quantifies the load distribution within a limb in a still or moving subject. Under this premise, the aim of this study was to test whether data derived from this parameter could detect the differences between sound and lame limbs in unilaterally lame dogs with elbow dysplasia. To accomplish this purpose, ten unilaterally lame dogs of similar conformation were walked over a pressure platform. Next, the COP path, in relation to the position of sound and lame limbs, was measured in a coordinate system over a standard paw template obtained by pedobarography during the whole support phase. To compare variables, force platform data (peak vertical force and vertical impulse) from the same animals were obtained. Sound and lame limb statokinesiograms were also obtained while the animals stood still. RESULTS: The statistical analysis clearly showed that COP in lame limbs start cranially and were shorter than sound limbs. In addition, the value of the COP excursion index was lower in lame limbs. Finally, the area of statokinesiograms was greater in lame limbs. CONCLUSION: This methodology based in limb COP characteristics serves to discriminate between sound and lame limbs in dogs with elbow dysplasia.

Assessment of the Efficacy of Platelet-Rich Plasma in the Treatment of Traumatic Canine Fractures.

The role of platelet-rich plasma (PRP) in promoting the healing of bone fractures has not yet been clearly stated. The aim of this prospective clinical study was to evaluate the effectiveness of plasma rich in growth factors (PRGF, a PRP derivate) in the treatment of naturally-occurring bone fractures in dogs. With this objective, sixty-five dogs with radius/ulna or tibia/fibula bone fractures were randomly divided into two groups (PRGF and saline solution (SS) groups) and checked at days 0, 7, 14, 21, 28, 35, 42, 49, 56, 60, 63, 70, 120, and 180. All the fractures were treated with an external skeletal fixation, and pain was controlled with Carprofen. Healing was evaluated by physical examination, limb function, radiography, and by a Likert-type owner satisfaction questionnaire. A faster fracture healing was observed in the PRGF group, with statistically significant differences with respect to the SS group. Swelling at the fracture site was significantly greater at day 14 and 28 in animals injected with PRGF, and more pain on palpation was found in the area at day 28. The injection of PRGF in acute bone fractures accelerates bone healing.

Adipose-Derived Mesenchymal Stem Cells: Are They a Good Therapeutic Strategy for Osteoarthritis?

Osteoarthritis (OA) is a major cause of disability in elderly population around the world. More than one-third of people over 65 years old shows either clinical or radiological evidence of OA. There is no effective treatment for this degenerative disease, due to the limited capacity for spontaneous cartilage regeneration. Regarding the use of regenerative therapies, it has been reported that one option to restore degenerated cartilage are adipose-derived mesenchymal stem cells (ASCs). The purpose of this review is to describe and compare the efficacy of ASCs versus other therapies in OA. METHODS: Recent studies have shown that ASCs exert paracrine effects protecting against degenerative changes in chondrocytes. According to the above, we have carried out a review of the literature using a combination of osteoarthritis, stem cells, and regenerative therapies as keywords. RESULTS: Conventional pharmacological therapies for OA treatment are considered before the surgical option, however, they do not stop the progression of the disease. Moreover, total joint replacement is not recommended for patients under 55 years, and high tibia osteotomy (HTO) is a viable solution to address lower limb malalignment with concomitant OA, but some complications have been described. In recent years, the use of mesenchymal stem cells (MSCs) as a treatment strategy for OA is increasing considerably, thanks to their capacity to improve symptoms together with joint functionality and, therefore, the patients' quality of life. CONCLUSIONS: ASC therapy has a positive effect on patients with OA, although there is limited evidence and little long-term follow-up.

Effect of autologous platelet-rich plasma on the repair of full-thickness articular defects in rabbits.

PURPOSE: To assess the effect of autologous platelet-rich plasma on the repair of full-thickness articular cartilage lesions in immature rabbits. To that end, the samples were studied using macroscopic, microscopic and biomechanical techniques. METHODS: Twenty-four 11-week-old New Zealand rabbits were divided into two groups based on the treatment (physiological saline serum PCB, autologous platelet-rich plasma PRP). Cartilage lesions were drilled in medial femoral condyle of these rabbits. Other 12 rabbits were under the same conditions, but they did not suffer any lesion and treatment (control group CTR). The three groups were divided into two subgroups, depending on the age at the time of killing (16 and 19 weeks old). RESULTS: The CTR group showed the best possible value in the macroscopic and microscopic evaluation. Meanwhile, the PCB and PRP group values were lower than the CTR group values, at two times of the study, but similar to each other at 19 weeks. In the biomechanical study, at 16 weeks, the CTR and PCB groups behaved similarly, with values above PRP group values, while at 19 weeks, CTR group showed higher values than PCB and PRP groups, and there were no differences between these values. CONCLUSIONS: The evolution of the tissue treated with autologous PRP showed a positive tendency over time, while the PCB group was negative. Nevertheless, at 19 weeks of age, the PRP treatment did not show better results than the PCB, both showing the characteristics of fibrocartilaginous tissue. Likewise, none of the two treatments produced a repair tissue as the healthy cartilage.

Experimental use of polyamide bands in combination with intramedullary pinning for repair of oblique femoral fractures in rabbits.

OBJECTIVE: To evaluate the use of polyamide bands, manufactured for securing electrical cables, for repair of oblique femoral fractures in rabbits. STUDY DESIGN: Experimental study. ANIMALS: Twelve nulliparous, 21-25-week-old, California female rabbits. METHODS: Rabbits were divided into 3 groups (n=4) and studied for 14, 28, or 56 days. A Z-shaped mid-diaphyseal femoral osteotomy was repaired with a 2.5 mm intramedullary pin and a polyamide 6.6 cerclage band. Healing was evaluated at intervals by physical examination, limb use, femoral radiographs, and callus histology. RESULTS: Rabbits had early limb use with good wound healing. From the 1st day, movement of the hip and stifle joints was satisfactory. Radiographically, fractures were healed at 28 days. Histologically, there was no foreign body reaction and bone healing was normal. CONCLUSION: Nylon cerclage band application was accomplished easily, maintained reduction, and resulted in good healing and limb use. CLINICAL RELEVANCE: Sterilized nylon bands, manufactured for electrical use, can be used for cerclage in rabbits.