Platelet rich plasma (PRP) enhances anabolic gene expression patterns in flexor digitorum superficialis tendons

Abstract

Platelet rich plasma (PRP) has recently been investigated for use in tissue regeneration studies that seek to utilize the numerous growth factors released from platelet a-granules. This study examined gene expression patterns, DNA, and collagen content of equine flexor digitorum superficialis tendon (SDFT) explants cultured in media consisting of PRP and other blood products. Blood and bone marrow aspirate (BMA) were collected from horses and processed to obtain plasma, PRP, and platelet poor plasma (PPP). IGF-I, TGF-b1, and PDGF-BB were quantified in all blood products using ELISA. Tendons were cultured in explant fashion with blood, plasma, PRP, PPP, or BMA at concentrations of 100%, 50%, or 10% in serum-free DMEM with amino acids. Quantitative RT-PCR for expression of collagen type I (COL1A1), collagen type III (COL3A1), cartilage oligomeric matrix protein (COMP), decorin, matrix metalloproteinase-3 (MMP-3), and matrix metalloproteinase-13 (MMP-13) was performed as were DNA and total soluble collagen assays. TGF-b1 and PDGF-BB concentrations were higher in PRP compared to all other blood products tested. Tendons cultured in 100% PRP showed enhanced gene expression of the matrix molecules COL1A1, COL3A1, and COMP with no concomitant increase in the catabolic molecules MMP-3 and MMP-13. These findings support in vivo investigation of PRP as an autogenous, patient-side treatment for tendonitis. ß

Ultrasound-guided injection of platelet-rich plasma in chronic Achilles and patellar tendinopathy

“…The significantly improved VISA scores at the 6-month follow-up were consistent with results previously obtained by other authors [38,39,46]. Also, the fact that tendon thickness and hypoechoic areas were reduced may be a sign of tendon regeneration, as collagen fibers were more closely packed like in normal tendons.…”

Mesenchymal stem cells and insulin‐like growth factor‐I gene‐enhanced mesenchymal stem cells improve structural aspects of healing in equine flexor digitorum superficialis tendons

“…Total cellular RNA was isolated using Trizol (Invitrogen, Carlsbad, CA) according to the manufacturer's directions with minor modifications as previously described, 24 and precipitated RNA was further purified using RNeasy spin columns (Qiagen, Chatsworth, CA). RNA was assessed by spectrophotometry at 260:280 nm and by 1% agarose gel electrophoresis.…”

“…RNA was assessed by spectrophotometry at 260:280 nm and by 1% agarose gel electrophoresis. RNA was used for quantitative RT/PCR of collagen types I and III, 22,24 IGF-I, 25 and cartilage oligomeric matrix protein (COMP), [26][27][28] as indicators of tendon matrix synthesis, and matrix metalloproteinase-3 (MMP-3), [29][30][31] matrix metalloproteinase-13 (MMP-13), 24,29,32 and aggrecanase (ADAMTS-4) 30,33 as indicators of tendon catabolism.…”

“…37 Total soluble collagen content was determined in lyophilized tendon samples using the Sircol Assay (Biocolor Ltd., Newtownabbey, Northern Ireland) according to the manufacturer's directions for pepsin soluble collagens with modifications as previously described. 24 Histology Paraformaldehyde-fixed longitudinal tissue sections for histology were dehydrated, cleared in xylene, embedded in paraffin, sectioned, mounted on microscope slides, and stained with hematoxylin and eosin (H&E). Sections for collagen type I immunohistochemistry were deparaffinized, rehydrated, and treated with 5 mL/mL hyaluronidase (Sigma-Aldrich, St. Louis, MO) at 378C for 60 min to facilitate antibody penetration into the tissue.…”

The state of cartilage regeneration: current and future technologies

“…As the cellular component includes platelets and MSCs, growth factors secreted include PDGF, TGF-B, and VEGF [34,35]. Using the Harvest system (Harvest Technologies, Plymouth, Massachusetts), significant increases in platelet, leukocytes, and red Using this system in an equine model, treatment of an osteochondral defect with microfracture and BM-MSCs demonstrated increased defect fill and type II collagen compared to microfracture alone [36••].…”