BACKGROUND: Recent evidence from in vitro and in vivo studies indicates that bisphosphonates may promote osteoblastic bone formation and potently inhibit osteoclast activity. However, little is known about the potential effect of bisphosphonates on the recruitment of osteoblastic precursors from patient-derived bone marrow stromal cells due to difficulties in accessing human bone marrow from healthy and disease subjects. METHODS: In this study, we evaluated the potential of using FDA-approved and clinically utilized bisphosphonates such as alendronate, ibandronate, and zoledronate to enhance the development of bone forming osteoblasts from osteoporosis patient- and healthy-person derived hBMSCs (op-MSCs and hp-MSCs, respectively). hBMSCs were obtained from postmenopausal women without endocrine diseases or receiving hormone replacement therapy. Cells were treated with or without a bisphosphonate (alendronate, ibandronate, and zoledronate) and analyzed over 21 days of culture. RESULTS: hBMSC from osteoporosis-patient with bisphosphonates treatment demonstrated a significant increase in Alizarin red staining after 7 days compared to that from healthy-person. Calcium contents and alkaline phosphatase (ALP) enzyme activity also demonstrated an increased propensity in hMSCs from osteoporosis-patient compared to those from healthy-person, although there were inter-individual variations.Gene expression levels varied among different donors. There were no significant differences in the effect on the osteoblastic differentiation of hBMSCs among alendronate, ibandronate, and zoledronate. Statistical significance in the osteoblastic differentiation of hBMSCs between the positive control group cultured in osteogenic mediumalone and groups cultured in osteogenic mediumsupplemented with bisphosphonate was not shown either.These results might be due to various cell types of hBMSCs from individual clinical patients and concentrations of bisphosphonate used. CONCLUSION: Our study using a clinically relevant in vitro model suggests that bisphosphonate treatment is more effective for patients with osteoporosis than its preventive effect for healthy person. In addition, patient-specific responses to bisphosphonates should be considered rather than bisphosphonate type prior to prescription. Further investigations are needed to determine how bisphosphonates influence hBMSCs function to mediate bone quality and turnover in osteoporotic patients. Such studies can generate novel approaches to treat age-related osteoporotic bone loss.
Alendronate ; Alkaline Phosphatase ; Bone Marrow ; Calcium ; Diphosphonates* ; Endocrine System Diseases ; Female ; Hormone Replacement Therapy ; Humans ; In Vitro Techniques ; Mesenchymal Stromal Cells* ; Osteoblasts ; Osteoclasts ; Osteogenesis ; Osteoporosis* ; Prescriptions ; Tissue Donors

BACKGROUND: Three-dimensional (3D) in vitro cultures recapitulate the physiological microenvironment and exhibit high concordance with in vivo conditions. Improving co-culture models with different kind of cell types cultured on a 3D scaffold can closely mimic the in vivo environment. In this study, we examined the osteogenic response of pre-osteoblast MC3T3-E1 cells and Raw264.7 mouse monocytes in a 3D-encapsulated co-culture environment composed of the Cellrix®3D culture system, which provides a physiologically relevant environment. METHODS: The Cellrix® 3D Bio-Gel scaffolds were used to individually culture or co-culture two type cells in 3D microenvironment. Under 3D culture conditions, osteoblastic behavior was evaluated with an ALP assay and staining. ACP assay and TRAP staining were used as osteoclastic behavior indicator. RESULTS: Treatment with osteoblastic induction factors (?3F) and RANKL had on positively effect on alkaline phosphatase activity but significantly inhibited to acid phosphatase activity during osteoclastic differentiation in 3D coculture. Interestingly, alkaline phosphatase activity or acid phosphatase activity in 3D co-culture was stimulated with opposite differentiation factors at an early stage of differentiation. We guess that these effects may be related to RANK– RANKL signaling, which is important in osteoblast regulation of osteoclasts. CONCLUSION In this study, the osteogenic response of 3D encapsulated pre-osteoblast MC3T3-E1 cells and mouse monocyte Raw264.7 cells was successfully demonstrated. Our 3D culture conditions will be able to provide a foundation for developing a high-throughput in vitro bone model to study the effects of various drugs and other agents on molecular pathways.

BACKGROUND: Three-dimensional (3D) in vitro cultures recapitulate the physiological microenvironment and exhibit high concordance with in vivo conditions. Improving co-culture models with different kind of cell types cultured on a 3D scaffold can closely mimic the in vivo environment. In this study, we examined the osteogenic response of pre-osteoblast MC3T3-E1 cells and Raw264.7 mouse monocytes in a 3D-encapsulated co-culture environment composed of the Cellrix®3D culture system, which provides a physiologically relevant environment. METHODS: The Cellrix® 3D Bio-Gel scaffolds were used to individually culture or co-culture two type cells in 3D microenvironment. Under 3D culture conditions, osteoblastic behavior was evaluated with an ALP assay and staining. ACP assay and TRAP staining were used as osteoclastic behavior indicator. RESULTS: Treatment with osteoblastic induction factors (?3F) and RANKL had on positively effect on alkaline phosphatase activity but significantly inhibited to acid phosphatase activity during osteoclastic differentiation in 3D coculture. Interestingly, alkaline phosphatase activity or acid phosphatase activity in 3D co-culture was stimulated with opposite differentiation factors at an early stage of differentiation. We guess that these effects may be related to RANK– RANKL signaling, which is important in osteoblast regulation of osteoclasts. CONCLUSION In this study, the osteogenic response of 3D encapsulated pre-osteoblast MC3T3-E1 cells and mouse monocyte Raw264.7 cells was successfully demonstrated. Our 3D culture conditions will be able to provide a foundation for developing a high-throughput in vitro bone model to study the effects of various drugs and other agents on molecular pathways.

PURPOSE: We investigated the effect of location changes in the inferior border of the belly board (BB) aperture by adding a bladder compression device (BCD). MATERIALS AND METHODS: We respectively reviewed data from 10 rectal cancer patients with a median age 64 years (range, 45~75) and who underwent computed tomography (CT) simulation with the use of BB to receive pelvic radiotherapy between May and September 2010. A CT simulation was again performed with the addition of BCD since small bowel (SB) within the irradiated volume limited boost irradiation of 5.4 Gy using the cone down technique after 45 Gy. The addition of BCD made the inferior border of BB move from symphysis pubis to the lumbosacral junction (LSJ). RESULTS: Following the addition of BCD, the irradiated volumes of SB and the abdominopelvic cavity (APC) significantly decreased (174.3+/-89.5 mL vs. 373.3+/-145.0 mL, p=0.001, 1282.6+/-218.7 mL vs. 1571.9+/-158 mL, p<0.001, respectively). Bladder volume within the treated volume increased with BCD (222.9+/-117.9 mL vs. 153.7+/-95.5 mL, p<0.001). The ratio of irradiated bladder volume to APC volume with BCD (33.5+/-14.7%) increased considerably compared to patients without a BCD (27.5+/-13.1%) (p<0.001), and the ratio of irradiated SB to APC volume decreased significantly with BCD (13.9+/-7.6% vs. 24.2+/-10.2%, p<0.001). The ratios of the irradiated SB volumeand irradiated bladder volume to APC volume negatively correlated (p=0.001). CONCLUSION: This study demonstrated that the addition of BCD, which made the inferior border of BB move up to the LSJ, increased the ratio of the bladder to APC volume and as a result, decreased the irradiated volume of SB.
Humans ; Rectal Neoplasms ; Urinary Bladder