Jong-pil Seo

Effects of bilayer gelatin/β-tricalcium phosphate sponges loaded with mesenchymal stem cells, chondrocytes, bone morphogenetic protein-2, and platelet rich plasma on osteochondral defects of the talus in horses.

Osteochondrosis (OC) is a common and clinically important joint disorder in horses. However, repair of the OC region is difficult because of the avascular nature of cartilage. This study aimed to evaluate the efficacy of bilayer gelatin/β-tricalcium phosphate (GT) sponges loaded with mesenchymal stem cells (MSCs), chondrocytes, bone morphogenetic protein-2 (BMP-2), and platelet rich plasma (PRP) for the repair of osteochondral defects of the talus in horses. Full-thickness osteochondral defects were created on both the lateral trochlear ridges of the talus (n = 6). In the test group, a basic GT sponge loaded with MSCs and BMP-2 (MSC/BMP2/GT) was inserted into the lower part of the defect, and an acidic GT sponge loaded with chondrocyte, MSCs, and PRP (Ch/MSC/PRP/GT) was inserted into the upper part of the defect. In the control group, the defect was treated only with bilayer GT sponges. Repair of osteochondral defects was assessed by radiography, quantitative computed tomography (QCT), and macroscopic and histological evaluation. The test group showed significantly higher radiographic, QCT, macroscopic, and histological scores than the control group. This study demonstrated that the bilayer scaffolds consisting of Ch/MSC/PRP/GT for the chondrogenic layer and MSC/BMP2/GT for the osteogenic layer promoted osteochondral regeneration in an equine model. The bilayer scaffolds described here may be useful for treating horses with OC.

In vivo osteoinductivity of gelatin β-tri-calcium phosphate sponge and bone morphogenetic protein-2 on an equine third metacarpal bone defect

This study evaluated the therapeutic effects of a gelatin-β-TCP sponge (sponge) incorporating BMP-2 (BMP-2/sponge) on bone regeneration in equines. Six bone defects were created in third metacarpals of five thoroughbred horses, and a total of six treatments were applied in a randomized manner. The treatments were BMP-2/sponge, BMP-2/gelatin hydrogel sheet (sheet), free BMP-2, bFGF/sheet, plain sponge, and plain sheet. The defects were monitored for 16 weeks by radiography and then examined by histological analysis. Radiographic evaluation scores of bone regeneration revealed significantly greater bone regeneration of defects treated with BMP-2/sponge than defects treated with plain sponge or BMP-2 sheet (P<0.05). In histological analysis, compact bone was observed over a wide area in the BMP-2/sponge treatment. We concluded that the treatment with BMP-2/sponge accelerated bone regeneration in the equines of this study.

Comparison of allogeneic platelet lysate and fetal bovine serum for in vitro expansion of equine bone marrow-derived mesenchymal stem cells

Mesenchymal stem cells (MSCs) are promising candidates for cell-based therapy and tissue engineering approaches. Fetal bovine serum (FBS) is commonly used for in vitro MSC expansion; however, the use of FBS may be associated with ethical, scientific, and safety issues. This study aimed to compare the ability of allogeneic platelet lysate (PL) and FBS to cause equine bone marrow-derived MSC expansion. MSCs were isolated from bone marrow aspirate in media supplemented with either PL or FBS, and cell proliferation properties and characteristics were examined. There were no significant differences in MSC yield, colony-forming unit-fibroblast (CFU-F) assay, and population doubling time between PL and FBS cultures. In addition, both PL-MSCs and FBS-MSCs showed similar results in term of ALP staining, osteogenic differentiation, and RT-PCR, although there were subtle differences in morphology, growth pattern, and adhesive properties. These results suggest that PL is a suitable alternative to FBS for use in equine MSC expansion, without the problems related to FBS use.

Proliferation of equine bone marrow-derived mesenchymal stem cells in gelatin/β-tricalcium phosphate sponges.

A three dimensional scaffold is essential in mesenchymal stem cells (MSCs) delivery in cell-based therapy for facilitating cell adherence, migration, proliferation, and differentiation. The objectives of this study were to evaluate the possibility of β-tricalcium phosphate incorporated gelatin sponges (Gelatin/β-TCP sponge) as scaffolds for equine MSCs and to examine the effects of seeding density and seeding method on the proliferation of equine MSCs in the Gelatin/β-TCP sponges. Mononuclear cells and MSCs isolated from bone marrow were seeded into Gelatin/β-TCP sponges at different densities by different seeding methods-static or agitated methods. Proliferation of the MSCs in Gelatin/β-TCP was assessed using the Cell Counting Kit-8 assay and histological examination. Distribution and proliferation of MSCs in the Gelatin/β-TCP sponge were observed, and the Gelatin/β-TCP sponge supported limited growth when seeded at high density. We also found that the agitated seeding method enhanced the proliferation of MSCs. This study demonstrated the suitability of Gelatin/β-TCP sponges for the proliferation and maintenance of equine MSCs. These results contribute to the application of MSC-seeded Gelatin/β-TCP sponges in equine medicine.

Bioengineered osteochondral precursor for treatment of osteochondritis dissecans in a Thoroughbred filly

CASE REPORT A 13-month-old Thoroughbred filly was diagnosed with osteochondritis dissecans (OCD) of the medial tibial malleolus. A sponge impregnated with platelet-rich plasma, bone morphogenetic protein-2, mesenchymal stem cells and gelatin β-tricalcium phosphate was applied to the OCD site following arthroscopy and debridement. Postoperative radiography (every week for 16 weeks), computed tomography (CT) (16 weeks postoperatively), arthroscopy (16 weeks postoperatively) and biopsy of the regenerated tissue (16 weeks postoperatively) were performed to evaluate the outcome. Radiographically, the defect began to diminish 3 weeks postoperatively and had disappeared by 12 weeks. CT images showed that the debrided site was filled with ossified tissue and arthroscopy showed that the regenerated tissue was covered with smooth tissue, which a biopsy showed was fibrocartilage. CONCLUSIONS Placing the impregnated sponge in the OCD lesion facilitated satisfactory regeneration of tissue in the debrided area, but the regenerated cartilage was fibrocartilage. This method may be a viable option for the treatment of cases of equine OCD, but further work to determine how to induce hyaline cartilage regeneration is required.

In vitro biomechanical comparison of a 5-hole 4.5 mm locking compression plate and 5-hole 4.5 mm dynamic compression plate for equine proximal interphalangeal joint arthrodesis.

OBJECTIVE To compare the biomechanical properties of a 5-hole 4.5 mm narrow locking compression plate (LCP) and 5-hole 4.5 mm narrow dynamic compression plate (DCP) for equine proximal interphalangeal (PIP) joint arthrodesis. STUDY DESIGN Experimental mechanical study. ANIMALS Cadaveric adult equine forelimbs (n = 6 pair). METHODS For each forelimb pair, 1 PIP joint was stabilized with LCP and the contralateral PIP joint with DCP. The 6 construct pairs were tested using a single-cycle, 3-point dorsopalmar bending system. PIP joints were evaluated with pre- and post-test radiography. RESULTS The LCP technique had significantly greater yield load, failure load, and stiffness under single-cycle, 3-point dorsopalmar bending to failure than the DCP technique. There was no significant difference between the 2 constructs for displacement at yield and failure point. CONCLUSIONS Biomechanically, the LCP technique provided significantly greater stability than the DCP technique under the test condition.Objective To compare the biomechanical properties of a 5-hole 4.5 mm narrow locking compression plate (LCP) and 5-hole 4.5 mm narrow dynamic compression plate (DCP) for equine proximal interphalangeal (PIP) joint arthrodesis. Study Design Experimental mechanical study. Animals Cadaveric adult equine forelimbs (n = 6 pair). Methods For each forelimb pair, 1 PIP joint was stabilized with LCP and the contralateral PIP joint with DCP. The 6 construct pairs were tested using a single-cycle, 3-point dorsopalmar bending system. PIP joints were evaluated with pre- and post-test radiography. Results The LCP technique had significantly greater yield load, failure load, and stiffness under single-cycle, 3-point dorsopalmar bending to failure than the DCP technique. There was no significant difference between the 2 constructs for displacement at yield and failure point. Conclusions Biomechanically, the LCP technique provided significantly greater stability than the DCP technique under the test condition.