Introduction
Recently, identification of the tendon derived stem cells(TDSCs) suggests a new paradigm to develop a new treatment of tendon healing and provision of adequate environment to differentiate these stem cells into normal tenocytes becomes important issue. Also bone marrow mesenchymal stem cells(BMMSCs) are good cell sources to repair the damaged structures and have been used to enhance the healing of the rotator cuff tear(RCT) in preclinical and clinical studies. However, without adequate scaffold, the survivorship, reparative and differentiation capacities of BMMSCs are not enough to enhance the healing of RCT. As a scaffold, Platelet rich plasma(PRP) is known to enhance the proliferation and multipotency of BMMSC. The purpose of this study is to investigate the effect of BMMSC with PRP on TDSC and RCT.
Methods
A. In-vitro study
TDSC was acquired from one patient with lateral epicondylopathy by tendon tissue biopsy. Characteristics of TDSC was indentified by Immunocytochemical staining and Fluorescence-activated cell sorting (FACS) analysis. And then difference between TDSC(alone) and TDSC-BMMSC-PRP(mixed) was investigated about Cell doubling time, differentiation assay, wound healing assay, Expression of collagen type I and III.
B. Clinical Study
8 adults enrolled in this study. After acquiring BMMSC and PRP from individual bone marrow and peripheral blood, 1 ml of BMMSC was mixed with 1 ml of PRP. They received the BMMSC-PRP injection at the tear site under the ultrasound guidance. Tear size, numeric rating scale (NRS) of pain were measured and American Shoulder Elbow Surgeon (ASES) scores were recorded before, 3 weeks after, and 3 months after the injection
Results
A. in-vitro study
Immunocytochemical staining of tenomodulin proved the identification of tenogenic-lineage of TDSC and Nucleostemin, Oct4, and SSEA4 proved the stemness of TDSCs. In FACS analysis, over 96% of TDSC expressed mesenchymal specific markers, including CD90 and CD105. Meanwhile they were negative for hematopoietic stem cell markers CD34 and leukocyte marker CD45(fig 1). Adipogenic and chondrogenic potentials of TDSCs were increased but osteogenic potential of TDSCs was decreased when they were co-cultured with BMMSC-PRP. Doubling time of TDSC was shortened by co-culture with BMMSC-PRP. Superiorty in wound healing assay was estimated in TDSC with BMMSC-PRP(fig 2). In the TDSC with BMMSC-PRP group, expression of collagen type I was higher than in TDSCs group. However, there was no difference of collagen type III expression.
B. clinical study
Initial NRS was 5.5±2.07, which changed to 3.2±2.13 at 3 months after the BMMSC-PRP injection. ASES changed from 43.3±13.2 to 73.1±16.41 at 3 months after the injection. Initial tear size of rotator cuff tendon was 10.9±11.47 mm2 and then size reduced to 4.1±6.92mm2 at 3 month after injection
Conclusion:
BMMSC-PRP is considered as a new paradigm to treat tendinopathy by enhancing proliferation and migration of TDSC |
