Francesca Salamanna1, Simona Cepollaro2, Deyanira Contartese3, Gianluca Giavaresi3, Giovanni Barbanti Brodano4, Cristiana Griffoni4, Alessandro Gasbarrini4, Milena Fini3. 1. Laboratory of Biocompatibility, Technological Innovation and Advanced Therapy, Rizzoli RIT, IRCCS Rizzoli Orthopedic Institute, Bologna, Italy. 2. Department of Medical and Surgical Sciences, University of Bologna, Bologna, Italy. 3. Laboratory of Preclinical and Surgical Studies, IRCCS Rizzoli Orthopedic Institute, Bologna, Italy. 4. Department of Oncological and Degenerative Spine Surgery, IRCCS Rizzoli Orthopedic Institute, Bologna, Italy.
Abstract
STUDY DESIGN: Laboratory study. OBJECTIVE: Mesenchymal stem cells (MSCs) derived from whole bone marrow aspirate (BMA) and MSCs derived from density-gradient centrifugation were isolated from vertebral bodies and cultured under either hypoxic or normoxic conditions to evaluate their biological characteristics and HOX and TALE signature able to improve spinal surgery procedures. SUMMARY OF BACKGROUND DATA: The use of spinal fusion procedures has increased over the last decades; however, failed fusion still remains an important problem. Clinician and researchers focused their attention on the therapeutic potential of bone marrow MSCs and several methods for their isolation and cultivation have been developed. However, the best source and techniques are still debated. METHODS: MSCs morphology, surface markers, colony-forming-units, and three lineage differentiation through quantitative real-time PCR (qPCR) were evaluated. Additionally, gene expression analysis of HOX and TALE signatures during osteogenic differentiation were analyzed. RESULTS: Our study showed that MSCs derived from whole BMA were successfully isolated and when cultured under hypoxic condition presented greater proliferation, larger colonies, and differentiated onto osteogenic and chondrogenic lineage with greater ability, while adipogenic differentiation was less efficient. Results also revealed that MSCs, differently isolated and cultured, expressed different level of HOX and TALE signatures and that HOXB8 were up-regulated with greater efficiency in MSCs derived from whole BMA under hypoxia. CONCLUSION: Our data indicated that hypoxic preconditioning of MSCs derived from whole BMA exhibited more suitable biological characteristics and different level of HOX and TALE gene activation. We, therefore, concluded that vertebral body MSCs derived from whole BMA may provide alternative sources of MSCs for tissue engineering applications for spine surgery. LEVEL OF EVIDENCE: N/A.
STUDY DESIGN: Laboratory study. OBJECTIVE: Mesenchymal stem cells (MSCs) derived from whole bone marrow aspirate (BMA) and MSCs derived from density-gradient centrifugation were isolated from vertebral bodies and cultured under either hypoxic or normoxic conditions to evaluate their biological characteristics and HOX and TALE signature able to improve spinal surgery procedures. SUMMARY OF BACKGROUND DATA: The use of spinal fusion procedures has increased over the last decades; however, failed fusion still remains an important problem. Clinician and researchers focused their attention on the therapeutic potential of bone marrow MSCs and several methods for their isolation and cultivation have been developed. However, the best source and techniques are still debated. METHODS: MSCs morphology, surface markers, colony-forming-units, and three lineage differentiation through quantitative real-time PCR (qPCR) were evaluated. Additionally, gene expression analysis of HOX and TALE signatures during osteogenic differentiation were analyzed. RESULTS: Our study showed that MSCs derived from whole BMA were successfully isolated and when cultured under hypoxic condition presented greater proliferation, larger colonies, and differentiated onto osteogenic and chondrogenic lineage with greater ability, while adipogenic differentiation was less efficient. Results also revealed that MSCs, differently isolated and cultured, expressed different level of HOX and TALE signatures and that HOXB8 were up-regulated with greater efficiency in MSCs derived from whole BMA under hypoxia. CONCLUSION: Our data indicated that hypoxic preconditioning of MSCs derived from whole BMA exhibited more suitable biological characteristics and different level of HOX and TALE gene activation. We, therefore, concluded that vertebral body MSCs derived from whole BMA may provide alternative sources of MSCs for tissue engineering applications for spine surgery. LEVEL OF EVIDENCE: N/A.
Authors: Marta Camacho-Cardenosa; Alba Camacho-Cardenosa; Rafael Timón; Guillermo Olcina; Pablo Tomas-Carus; Javier Brazo-Sayavera Journal: Int J Environ Res Public Health Date: 2019-05-21 Impact factor: 3.390
Authors: F Salamanna; D Contartese; G Giavaresi; L Sicuro; G Barbanti Brodano; A Gasbarrini; M Fini Journal: Sci Rep Date: 2020-03-05 Impact factor: 4.379