RESUMEN
Porous tantalum (Ta) implants have been successful in various orthopedic procedures for patients with compromised bone-forming abilities. Previous studies demonstrated that human osteoblast (HOB) cultures from older female patients produced less bone on implant materials in vitro compared to HOBs from age-matched male and younger female patients. In this study, the responses of HOBs from younger (< 45) and older (> 60 years old) female patients were compared on Ta, titanium fiber mesh (TFM) and tissue culture plastic. Adhesion, proliferation, and mineralization were greater in cells from younger patients than from older patients. Cell adhesion was slightly higher on Ta than TFM or plastic. However, Ta highly stimulated cell proliferation with a 4- and 6-fold increase compared to TFM for cells from younger and older patients, respectively, and 12- and 16-fold increase in proliferation compared to cells on plastic (p ≤ 0.001). At 3 weeks, mineralization was significantly higher on Ta compared to TFM for HOBs from older patients (p ≤ 0.05). Expression levels of bone matrix markers demonstrated differences dependent on age and substrate. Scanning electron micrographs revealed HOBs covering the surfaces and entering the pores of both Ta and TFM. In conclusion, tantalum greatly stimulates cell proliferation, and improves the ability of HOBs from older patients to form bone.
Asunto(s)
Materiales Biocompatibles/farmacología , Osteoblastos/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Tantalio/farmacología , Titanio/farmacología , Adulto , Anciano , Biomarcadores/metabolismo , Calcinosis/inducido químicamente , Calcinosis/metabolismo , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Femenino , Humanos , Persona de Mediana Edad , Osteoblastos/metabolismo , Osteoblastos/ultraestructura , Osteogénesis/fisiología , Adulto JovenRESUMEN
Complementary and alternative medicine (CAM) techniques are commonly used in hospitals and private medical facilities; however, the effectiveness of many of these practices has not been thoroughly studied in a scientific manner. Developed by Dr. Dolores Krieger and Dora Kunz, Therapeutic Touch is one of these CAM practices and is a highly disciplined five-step process by which a practitioner can generate energy through their hands to promote healing. There are numerous clinical studies on the effects of TT but few in vitro studies. Our purpose was to determine if Therapeutic Touch had any effect on osteoblast proliferation, differentiation, and mineralization in vitro. TT was performed twice a week for 10 min each on human osteoblasts (HOBs) and on an osteosarcoma-derived cell line, SaOs-2. No significant differences were found in DNA synthesis, assayed by [(3)H]-thymidine incorporation at 1 or 2 weeks for SaOs-2 or 1 week for HOBs. However, after four TT treatments in 2 weeks, TT significantly (p = 0.03) increased HOB DNA synthesis compared to controls. Immunocytochemistry for Proliferating Cell Nuclear Antigen (PCNA) confirmed these data. At 2 weeks in differentiation medium, TT significantly increased mineralization in HOBs (p = 0.016) and decreased mineralization in SaOs-2 (p = 0.0007), compared to controls. Additionally, Northern blot analysis indicated a TT-induced increase in mRNA expression for Type I collagen, bone sialoprotein, and alkaline phosphatase in HOBs and a decrease of these bone markers in SaOs-2 cells. In conclusion, Therapeutic Touch appears to increase human osteoblast DNA synthesis, differentiation and mineralization, and decrease differentiation and mineralization in a human osteosarcoma-derived cell line.
Asunto(s)
Calcificación Fisiológica/fisiología , ADN/biosíntesis , Osteoblastos/citología , Osteosarcoma/patología , Tacto Terapéutico , Biomarcadores/metabolismo , Línea Celular Tumoral , Proliferación Celular , Regulación de la Expresión Génica , Humanos , Persona de Mediana Edad , Osteoblastos/metabolismo , Osteosarcoma/genética , Osteosarcoma/metabolismo , Antígeno Nuclear de Célula en Proliferación/metabolismo , ARN Mensajero/metabolismoRESUMEN
OBJECTIVES: Our objective was to assess the effect of Therapeutic Touch (TT) on the proliferation of normal human cells in culture compared to sham and no treatment. Several proliferation techniques were used to confirm the results, and the effect of multiple 10-minute TT treatments was studied. DESIGN: Fibroblasts, tendon cells (tenocytes), and bone cells (osteoblasts) were treated with TT, sham, or untreated for 2 weeks, and then assessed for [(3)H]-thymidine incorporation into the DNA, and immunocytochemical staining for proliferating cell nuclear antigen (PCNA). The number of PCNA-stained cells was also quantified. For 1 and 2 weeks, varying numbers of 10-minute TT treatments were administered to each cell type to determine whether there was a dose-dependent effect. RESULTS: TT administered twice a week for 2 weeks significantly stimulated proliferation of fibroblasts, tenocytes, and osteoblasts in culture (p = 0.04, 0.01, and 0.01, respectively) compared to untreated control. These data were confirmed by PCNA immunocytochemistry. In the same experiments, sham healer treatment was not significantly different from the untreated cultures in any group, and was significantly less than TT treatment in fibroblast and tenocyte cultures. In 1-week studies involving the administration of multiple 10-minute TT treatments, four and five applications significantly increased [(3)H]-thymidine incorporation in fibroblasts and tenocytes, respectively, but not in osteoblasts. With different doses of TT for 2 weeks, two 10-minute TT treatments per week significantly stimulated proliferation in all cell types. Osteoblasts also responded to four treatments per week with a significant increase in proliferation. Additional TT treatments (five per week for 2 weeks) were not effective in eliciting increased proliferation compared to control in any cell type. CONCLUSIONS: A specific pattern of TT treatment produced a significant increase in proliferation of fibro-blasts, osteoblasts, and tenocytes in culture. Therefore, TT may affect normal cells by stimulating cell proliferation.
Asunto(s)
Fibroblastos/citología , Fibroblastos/metabolismo , Osteoblastos/citología , Osteoblastos/metabolismo , Tacto Terapéutico/métodos , Proliferación Celular , Células Cultivadas , Humanos , Inmunohistoquímica , Antígeno Nuclear de Célula en Proliferación/metabolismo , Coloración y Etiquetado , Timidina/metabolismo , Timidina/fisiologíaRESUMEN
Tendon to bone integration after rotator cuff repair is not a reproducible process. During repair, bioabsorbable and nonabsorbable suture material is universally used to facilitate the procedure. Improving the biological architecture of inert suture might aid in overall tendon to bone healing. The objective of our study is to enhance the bone to tendon union by absorbing type I collagen onto high strength nonabsorbable polyester/polyethylene suture commonly used in rotator cuff surgery. Our purpose was to evaluate the tendon and bone cellular response to this novel coated suture compared to uncoated suture. Primary human osteoblasts (HOBs) and tenocytes were plated onto polyester/polyethylene suture that was either uncoated or coated with type I bovine collagen. Cell adhesion to the sutures was assayed at 24 hours. Proliferation was determined at 48 hours by measuring [3H]- Thymidine incorporation in cells attached to the sutures. At 24 and 48 hours, respectively, cells grown on the collagen-coated suture showed a significantly greater response measured by adhesion and proliferation than cells grown on uncoated suture. At five days of culture, alkaline phosphatase activity and protein synthesis was significantly greater on the collagen-coated suture compared to uncoated. Collagen-coated polyester/polyethylene suture appears to stimulate adhesion, proliferation alkaline phosphatase, and protein synthesis more than uncoated sutures, and therefore may aid in the tendon to bone incorporation process critical to rotator cuff repair.