In Vivo Evaluation of Porous Hydroxyapatite/Poly D/L-Lactide Composite for Bone Substitute and Scaffold

2005 ◽  
Vol 284-286 ◽  
pp. 769-774
Author(s):  
Shin Hasegawa ◽  
Jiro Tamura ◽  
Masashi Neo ◽  
Koji Goto ◽  
Yasuo Shikinami ◽  
...  

We investigated the biocompatibility, osteoconductivity, and biodegradability of porous composite of Hydroxyapatite (HA) and Poly D/L-lactide (PDLLA). At 6weeks afterimplantation to rabbit femoral condyle, HA/PDLLA was covered with bone and contacted with bone directly. The amounts of newly formed bone in the pores had increased during the examined period. By 26weeks, bone remodeling of formed bone in the pores was seen and bone marrow tissue formation was seen in the pores of HA/PDLLA. Porous HA/PDLLA was resorbed much faster than porous HA as a control. Porous HA/PDLLA was resorbed constantly through the bone formation and bone remodeling but porous HA was hardly resorbed during the period. It might be one of the desirable materials for bone substitute. To evaluate for a scaffold, disc shaped blocks loaded with rat bone marrow cell were implanted in the subcutaneous pouch of the back of syngeneic rat. At 3weeks afterimplantation, newly bone formation in the pores was observed at ectopic site. It also suggested the availability of this material as cell scaffolds.

Author(s):  
Shin Hasegawa ◽  
Jiro Tamura ◽  
Masashi Neo ◽  
Koji Goto ◽  
Yasuo Shikinami ◽  
...  

2007 ◽  
Vol 361-363 ◽  
pp. 1189-1192 ◽  
Author(s):  
N. Tsuji ◽  
Masataka Yoshikawa ◽  
Y. Shimomura ◽  
T. Yabuuchi ◽  
H. Hayashi ◽  
...  

The purpose of this study was to estimate influence of lysine for osteogenesis in the porous hydroxyapatite (HA) scaffolds with bone marrow cells. The HA scaffolds were soaked in 100mM concentration of lysine solution. They were kept in bone marrow cell suspension at 1×106 cells/ml density. Another HA scaffolds without immersion in lysine solution were kept in the cell suspension at 1×106 or 1×107 cells/ml density. They were respectively implanted into dorsal subcutis of rats for 4 weeks. Serially sectioned paraffin specimens were made and observed histologically. In several sections, total pores and ones with bone were counted. Many pores containing bone were found in1×107 cells/ml concentration group. The significant difference was between 1×107 cells/ml group, the lysine group, and 1×106 cells/ml group. Although more bone formation was seen in lysine group than in 1×106 cells/ml group. There was no significant difference between the groups. Concentration of lysine to add in culture medium or scaffold should be improved respectively.


2005 ◽  
Vol 75A (3) ◽  
pp. 567-579 ◽  
Author(s):  
Shin Hasegawa ◽  
Jiro Tamura ◽  
Masashi Neo ◽  
Koji Goto ◽  
Yasuo Shikinami ◽  
...  

2021 ◽  
Vol 12 (7) ◽  
Author(s):  
Chao Liu ◽  
An-Song Liu ◽  
Da Zhong ◽  
Cheng-Gong Wang ◽  
Mi Yu ◽  
...  

AbstractBone marrow-derived mesenchymal stem cells (BM-MSCs), the common progenitor cells of adipocytes and osteoblasts, have been recognized as the key mediator during bone formation. Herein, our study aim to investigate molecular mechanisms underlying circular RNA (circRNA) AFF4 (circ_AFF4)-regulated BM-MSCs osteogenesis. BM-MSCs were characterized by FACS, ARS, and ALP staining. Expression patterns of circ_AFF4, miR-135a-5p, FNDC5/Irisin, SMAD1/5, and osteogenesis markers, including ALP, BMP4, RUNX2, Spp1, and Colla1 were detected by qRT-PCR, western blot, or immunofluorescence staining, respectively. Interactions between circ_AFF4 and miR-135a-5p, FNDC5, and miR-135a-5p were analyzed using web tools including TargetScan, miRanda, and miRDB, and further confirmed by luciferase reporter assay and RNA pull-down. Complex formation between Irisin and Integrin αV was verified by Co-immunoprecipitation. To further verify the functional role of circ_AFF4 in vivo during bone formation, we conducted animal experiments harboring circ_AFF4 knockdown, and born samples were evaluated by immunohistochemistry, hematoxylin and eosin, and Masson staining. Circ_AFF4 was upregulated upon osteogenic differentiation induction in BM-MSCs, and miR-135a-5p expression declined as differentiation proceeds. Circ_AFF4 knockdown significantly inhibited osteogenesis potential in BM-MSCs. Circ_AFF4 stimulated FNDC5/Irisin expression through complementary binding to its downstream target molecule miR-135a-5p. Irisin formed an intermolecular complex with Integrin αV and activated the SMAD1/5 pathway during osteogenic differentiation. Our work revealed that circ_AFF4, acting as a sponge of miR-135a-5p, triggers the promotion of FNDC5/Irisin via activating the SMAD1/5 pathway to induce osteogenic differentiation in BM-MSCs. These findings gained a deeper insight into the circRNA-miRNA regulatory system in the bone marrow microenvironment and may improve our understanding of bone formation-related diseases at physiological and pathological levels.


Blood ◽  
1996 ◽  
Vol 87 (10) ◽  
pp. 4136-4142 ◽  
Author(s):  
I Kawashima ◽  
ED Zanjani ◽  
G Almaida-Porada ◽  
AW Flake ◽  
H Zeng ◽  
...  

Using in utero transplantation into fetal sheep, we examined the capability of human bone marrow CD34+ cells fractionated based on Kit protein expression to provide long-term in vivo engraftment. Twelve hundred to 5,000 CD34+ Kit-, CD34+ Kit(low), and CD34+ Kit(high) cells were injected into a total of 14 preimmune fetal sheep recipients using the amniotic bubble technique. Six fetuses were killed in utero 1.5 months after bone marrow cell transplantation. Two fetuses receiving CD34+ Kit(low) cells showed signs of engraftment according to analysis of CD45+ cells in their bone marrow cells and karyotype studies of the colonies grown in methylcellulose culture. In contrast, two fetuses receiving CD34+ Kit(high) cells and two fetuses receiving CD34+ Kit- cells failed to show evidence of significant engraftment. Two fetuses were absorbed. A total of six fetuses receiving different cell populations were allowed to proceed to term, and the newborn sheep were serially examined for the presence of chimerism. Again, only the two sheep receiving CD34+ Kit(low) cells exhibited signs of engraftment upon serial examination. Earlier in studies of murine hematopoiesis, we have shown stage-specific changes in Kit expression by the progenitors. The studies of human cells reported here are in agreement with observations in mice, and indicate that human hematopoietic stem cells are enriched in the Kit(low) population.


Blood ◽  
1993 ◽  
Vol 82 (1) ◽  
pp. 318-325 ◽  
Author(s):  
NJ Parks ◽  
TG Kawakami ◽  
MJ Avila ◽  
R White ◽  
GR Cain ◽  
...  

beta-emitting 166Ho (t1/2 = 26.78 hours, E(beta)max = 1.8 MeV) complexed with the phosphonic acid chelator, 1,4,7,10 tetraazacyclododecane-1,4,7,10-tetra(methylene phosphonic acid) (DOTMP) at a ligand-to-metal ratio of 1.5:1 binds to bone. This radioactive complex is a marrow-ablating radiopharmaceutical that appears useful for preparation of bone marrow (BM) transplant recipients without the morbidity usually associated with total body irradiation preparatory regimens. We have found with seven splenectomized young adult beagle dogs that a 166Ho radiopharmaceutical dosage of 370 MBq/kg body weight provides an initial skeletal radioactivity burden of at least 1.5 GBq/kg skeleton and results in complete ablation of hematopoietic marrow cell populations within 7 days. The beta particle flux distribution in BM-forming skeletal tissue is not uniform. Red marrow radiation doses varied from 30 to 115 Gy as estimated by direct radioassay and autoradiographic analyses of both bone biopsies and postmortem samples; the median value of 61 Gy agreed with our theoretical expectations. In vivo radioactivity distribution was evaluated with nuclear imaging methods. Apparently, normal hematopoiesis was restored in three dogs with autologous BM transplants performed 5 to 6 days after administration of the marrow ablative radiopharmaceutical, 166Ho-DOTMP. BM biopsies at 7 to 10 months posttransplantation indicate continued normal hematopoietic activity.


2018 ◽  
Vol 16 (3) ◽  
pp. 126-136 ◽  
Author(s):  
Preeti Makkar ◽  
Swapan Kumar Sarkar ◽  
Andrew R. Padalhin ◽  
Byoung-Gi Moon ◽  
Young Seon Lee ◽  
...  

Background: Magnesium (Mg)-based alloys are considered to be promising materials for implant application due to their excellent biocompatibility, biodegradability, and mechanical properties close to bone. However, low corrosion resistance and fast degradation are limiting their application. Mg–Ca alloys have huge potential owing to a similar density to bone, good corrosion resistance, and as Mg is essential for Ca incorporation into bone. The objective of the present work is to determine the in vitro degradation and in vivo performance of binary Mg– xCa alloy ( x = 0.5 or 5.0 wt%) to assess its usability for degradable implant applications. Methods: Microstructural evolutions for Mg– xCa alloys were characterized by optical, SEM, EDX, and XRD. In vitro degradation tests were conducted via immersion test in phosphate buffer saline solution. In vivo performance in terms of interface, biocompatibility, and biodegradability of Mg– xCa alloys was examined by implanting samples into rabbit femoral condyle for 2 and 4 weeks. Results: Microstructural results showed the enhancement in intermetallic Mg2Ca phase with increase in Ca content. Immersion tests revealed that the dissolution rate varies linearly, with Ca content exhibiting more hydrogen gas evolution, increased pH, and higher degradation for Mg–5.0Ca alloy. In vivo studies showed good biocompatibility with enhanced bone formation for Mg–0.5Ca after 4 weeks of implantation compared with Mg–5.0Ca alloy. Higher initial corrosion rate with prolonged inflammation and rapid degradation was noticed in Mg–5.0Ca compared with Mg–0.5Ca alloy. Conclusions: The results suggest that Mg–0.5Ca alloy could be used as a temporary biodegradable implant material for clinical applications owing to its controlled in vivo degradation, reduced inflammation, and high bone-formation capability.


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