Histological observation of a gelatin sponge transplant loaded with bone marrow-derived mesenchymal stem cells combined with platelet-rich plasma in repairing an annulus defect.
OBJECTIVE
To research the histological characteristics of a gelatin sponge transplant loaded with goat BMSCs (bone marrow-derived mesenchymal stem cells) combined with PRP (platelet-rich plasma) in repairing an annulus defect.


METHODS
BMSCs were separated from the iliac crest of goats, sub-cultured and identified after the third generation. Then, PRP was obtained using blood from the jugular vein of goats via two degrees of centrifugation. In the animal experiments, the goats were divided into the following three groups: a sham group, an injury group and a therapeutic group. In the sham group, we decompressed the lamina and exposed the annulus fibrosus. In the injury group, we exposed the annulus fibrosus after decompression of the lamina and created a 1 × 1 cm defect in the annulus using surgical instruments. In the therapeutic group, after decompression of the lamina, we exposed the annulus, created a 1 × 1 cm defect using surgical instruments, and placed a gelatin sponge combined with BMSCs and PRP into the defect for a combined method of repair. Three, six and twelve weeks after the surgery, the previously damaged or undamaged annulus tissue was removed from the three groups. Then, the above tissue was assayed using HE (hematoxylin-eosin) staining, Masson trichrome staining, AB-PAS (Alcian blue-periodic acid Schiff) staining, and type II collagen staining and observed by microscopy.


RESULTS
From the HE staining, we observed that the number of repair cells gradually increased. Compared to the injury group, the cell density and gross morphology of cells in the therapeutic group were closer to those of the sham group. As observed by Masson trichrome gelatin staining, many of the fibroblast cells or tissues were under repair, and as time progressed, the number of fibroblast cells and amount of tissue gradually increased. The results of the AB-PAS staining suggest that chondrocytes participated in the repair of the annulus. The level of type II collagen gradually increased, as determined by immunohistochemical staining.


CONCLUSIONS
Our results demonstrate that a gelatin sponge transplant loaded with BMSCs and PRP can effectively repair annulus defects.
Department of Spine Surgery, Xiangya Hospital, Central South University, Changsha, PR China.
Xiang
X
Xu
Department of Spine Surgery, Xiangya Hospital, Central South University, Changsha, PR China.|Key Laboratory of Organ Injury, Aging and Regenerative Medicine of Hunan Province, Changsha, PR China.
Jianzhong
J
Hu
Key Laboratory of Organ Injury, Aging and Regenerative Medicine of Hunan Province, Changsha, PR China.|Department of Sports Medicine, Research Centre of Sports Medicine, Xiangya 10 Hospital, Central South University, Changsha, PR China.
Hongbin
H
Lu
D000818:Animals; D000070616:Annulus Fibrosus / Q000473:pathology*; D024043:Collagen Type II / Q000378:metabolism; D005780:Gelatin / Q000737:chemistry; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D023421:Models, Animal; D053657:Platelet-Rich Plasma; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry; D014945:Wound Healing
D024043:Collagen Type II; D005780:Gelatin