• Title/Summary/Keyword: MC3T3 cell

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EFFECTS OF SOME RESTORATIVE MATERIALS ON THE PROLIFERATION OF OSTEOBLASTIC CELL AND THE TISSUE REACTION OF BONE (수종 수복재의 조골세포 유사세포 증식 및 골조직 반응에 미치는 영향)

  • Kim, Hyun-Sun;Hong, Chan-Ui;Kim, Jong-Yeo
    • Restorative Dentistry and Endodontics
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    • v.22 no.1
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    • pp.305-324
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    • 1997
  • Numerous materials such as amalgam, IRM, SuperEBA, dessicated ZOE, and Ketac-Silver have been used as a root-end filling material or to repair furcation perforations. But so far no material has been found to satisfy all of the requirements of an ideal restorative material. Recently, mineral trioxide aggregate (MTA) has been suggested for use as a root end filling material and for the repair of furcation perforations. The purpose of this study was to compare the effect of MTA on the proliferation of MC3T3/E1 osteoblastic cell, formation of bone nodule, alkaline phosphatase activity, and finally the tissue reaction of bone with those of amalgam, IRM, SuperEBA, dessicated ZOE, and Ketac-Silver. The following conclusions were drawn within the limits of the experimental results : 1. MTA showed a excellent proliferation of osteoblastic cell and Ketac-Silver showed moderate proliferation of osteoblastic cell. The rest of test materials showed no proliferation of osteoblastic cell. 2. Many of definite bone nodules were found in the MTA group. In contrast, Ketac-Silver group showed no definite bone formation but only showed mild sign of bone formation. 3. Alkaline phosphatase activity of Ketac-Silver and MTA showed similar results. But both of them showed higher activity than that of other materials (p<0.005). 4. The tissue reaction to implanted MTA in the calbarium of mouse was milder than that observed with other materials. The tissue reaction of dessicated ZOE showed the worst results among the test materials.

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The Effect of Electrical Stimulation on Osteoblast Surrounding Dental Implant (치과용 임플란트 주위 뼈모세포에 대한 전기자극의 영향에 관한 연구)

  • Woo, Kyung-Yeup;Kwon, Kung-Rock;Choi, Boo-Byung
    • Journal of Dental Rehabilitation and Applied Science
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    • v.19 no.3
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    • pp.195-206
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    • 2003
  • Several factors can affect the formation of bone tissues surrounding implants. One of the factors is electrical stimulation. It is known to change the movement of cells, form and destroy cells, and alter concentration and chemical component of soft tissues and bones. The effect of electrical stimulation on bone formation can vary according to the intensity of electric currents, stimulating time, the method of sending electric currents, and tissues and cells currents are applied to. This study examines how various enviroments affect osteoblasts. (1) effect on osteoblast with varying intensity of currents Osteoblast-like cells were raised on four plates where implants can be placed. A constant current sink (MC3T3-E1) that can adjust the intensity and stimulating time of electric currents was used. The four plates were stimulated with $0{\mu}A$, $10{\mu}A$, $20{\mu}A$, and $40{\mu}A$, respectively. After 24 hours of stimulation, the number and distribution of cells surrounding implants were examined. (2) effect on osteoblast with varying conditions The 3 study was performed with same method. (1) The change of attached cell number 72-hour after application of various currents (2) The change of attached cell number 72-hour after application of various interval (3) The comparison of attached cell number by implant surface texture The following are the results: 1. The distribution and density of cells surrounding implant is highest under the intensity of electric currents of $20{\mu}A$. 2. The number of cells attached implants is highest under the intensity of electric currents of $20{\mu}A$. 3. The number of cells attached implants is highest under continous electric currents 4. The number of cells attached implants is not different by implant surface texture.

Microgrooves on titanium surface affect peri-implant cell adhesion and soft tissue sealing; an in vitro and in vivo study

  • Lee, Hyo-Jung;Lee, Jaden;Lee, Jung-Tae;Hong, Ji-Soo;Lim, Bum-Soon;Park, Hee-Jung;Kim, Young-Kwang;Kim, Tae-Il
    • Journal of Periodontal and Implant Science
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    • v.45 no.3
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    • pp.120-126
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    • 2015
  • Purpose: With the significance of stable adhesion of alveolar bone and peri-implant soft tissue on the surface of titanium for successful dental implantation procedure, the purpose of this study was to apply microgrooves on the titanium surface and investigate their effects on peri-implant cells and tissues. Methods: Three types of commercially pure titanium discs were prepared; machined-surface discs (A), sandblasted, large-grit, acid-etched (SLA)-treated discs (B), SLA and microgroove-formed discs (C). After surface topography of the discs was examined by confocal laser scanning electron microscopy, water contact angle and surface energy were measured. Human gingival fibroblasts (hGFs) and murine osteoblastic cells (MC3T3-E1) were seeded onto the titanium discs for immunofluorescence assay of adhesion proteins. Commercially pure titanium implants with microgrooves on the coronal microthreads design were inserted into the edentulous mandible of beagle dogs. After 2 weeks and 6 weeks of implant insertion, the animal subjects were euthanized to confirm peri-implant tissue healing pattern in histologic specimens. Results: Group C presented the lowest water contact angle ($62.89{\pm}5.66{\theta}$), highest surface energy ($45{\pm}1.2mN/m$), and highest surface roughness ($Ra=22.351{\pm}2.766{\mu}m$). The expression of adhesion molecules of hGFs and MC3T30E1 cells was prominent in group C. Titanium implants with microgrooves on the coronal portion showed firm adhesion to peri-implant soft tissue. Conclusions: Microgrooves on the titanium surface promoted the adhesion of gingival fibroblasts and osteoblastic cells, as well as favorable peri-implant soft tissue sealing.

Study of Mylabris Phalerata on Anti-cancer Effects in Some Kinds of Cancer Cells (반모가 수종의 인체 암세포에 미치는 영향)

  • Kim, Jin-Sung;Yoon, Sang-Hyub;Ryu, Bong-Ha;Ryu, Ki-Won;Jung, Myung-Chai
    • The Journal of Internal Korean Medicine
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    • v.25 no.2
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    • pp.202-213
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    • 2004
  • Object : Objective: This study was conducted to investigate the anti-cancer effects of Mylabris phalerata (반모) in some kinds of cancer cells. Materials and Methods: Some kinds of cancer cells lines were treated. We used nine kinds of cancer cell lines, such as stomach cancer cells (Kato), lung cancer cells (Calu-1, NCI-H 1395), urinary bladder cancer cells (HS789T), bone cancer cells (Saos-2), brain cancer cells (SK-N-MC), liver cancer cells (Hep-G2), skin cancer cells (Mo-1) and prostate cancer cells (PC-3) with the water decoction of Mylabris phalerata. The histological changes of all cell lines in the media (RPMI-1640) containing the decoction of Mylabris phalerata were observed and we examined cell death assay by trypan blue exclusion testing was examined. Finally, the change of mitochondrial membrane potential was measurd and the inhibitory effect of Mylabris phalerata on cell increase was examined by analyzing the cell cycle. Results: In histologic change all cancer cell lines showed withdrawn and floating appearance that is typical in cellular impairment. Most of the cell lines showed over 50% death rate after 24 hours in trypan blue exclusion tests. Especially the stomach, urinary bladder. brain and liver cell lines showed over 30% death rate after 12 hours. All cell lines treated with Mylabris phalerata were less stained than the control group and the mitochondrial membrane potential in the Mylabris phalerata treated cell lines was markedly lower than that in the control group. The measurement of DNA quantity in all cell lines showed the disappearance of the peak and the thickened left axis, which suggests that all cellular DNA degraded. Conclusion: Mylabris phalerata had cytotoxicity on various kinds of cancer cell lines and the mechanism of that was the impairment of mitochondria by the breakdown of the mitochondrial cell membrane. We propose that this is in part attributable to the destruction of DNA in cancer cells.

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The Effects of Prostaglandin and Dibutyryl cAMP on Osteoblastic Cell Activity and Osteoclast Generation (Prostaglandin과 Dibutyryl cAMP가 조골세포의 활성과 파골세포 형성에 미치는 영향)

  • Mok, Sung-Kyu;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.448-468
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    • 1996
  • To maintain its functional integrity, bone is continuously remodelled by a process involving resorption by osteoeclasts and formation by osteoblasts, In order to respond to changes in the physical environment or to trauma with the relevant action, this process is strictly regulated by locally synthesized or systemic fators, Prostaglandin $E_2(PGE_2$) is perhaps one of the best studied factors, having been known to affect bone cell function for several decades.$PGE_2$ has both anabolic and catabolic activities. Excess of $PGE_2$ has been implicated in a number of pathological states associated with bone loss in a number of chronic inflammatory conditions such as periodontal disease and rheumatoid arthritis. $PGE_2$ and other arachidonic acid metabolites have been shown to be potent stimulators of osteoclastic bone resorption in organ culture. The anabolic effects of $PGE_2$ were first noticed when an increase in periosteal woven bone formation was seen after the infusion of $PGE_2$ into infants in order to prevent closure of the ductus arteriosus. The cellular basis for the catabolic actions of $PGE_2$ has been well characterized. $PGE_2$increases osteoclast recruitment in bone marrow cell cultures. Also $PGE_2$ has a direct action on osteoclast serving to inhibit activity and can also indirectly activate osteoclast via other cells in the vicinity, presumably osteoblast. The cellular mechanisms for the anabolic actions of $PGE_2$ are not nearly so well understood. The purpose of this paper was to study the effects of $PGE_2$ and dibutyl(DB)cAMP on osteoblastic clone MC3T3El cells and on the generation of osteoclasts from their precursor cells. The effect of $PGE_2$ and DBcAMP on the induction of alkaline phoaphatase(AlP) was investigated in osteoblastic clone MC3T3El cells cultured in medium containing 0.4% fetal bovine serum. $PGE_2$ and DBcAMP stimulated ALP activity and MTT assay in the cells in a dose-dependent manner at concentrations of lO-SOOng/ml. Cycloheximide, protein synthesis inhibitor, inhibited the stimulative effect of $PGE_2$ and DBcAMP on ALP activity in the cells. $PGE_2$also increased the intracellular cAMP content in a dose-dependent fashion with a maximal effect at 500ng/ml. The effect of $PGE_2$ on the generation of osteoclasts was investigated in a coculture system of mouse bone marrow cells with primary osteoblastic cells cultured in media containing 10% fetal bovine serum.After cultures, staining for tartrate-resistant acid phosphatase(TRAP)-marker enzyme of osteoclast was performed. The TRAP(+) multinucleated cells(MNCs), which have 3 or more nuclei, were counted. More TRAP(+) MNCs were formed in coculture system than in control group. $PGE_2(10^{-5}10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in culture. $PGE_2(10^{-6}M)$ stimulated the formation of osteoclast cells from mouse bone marrow cells in coculture of osteoblastic clone MC3T3E1 cells This results suggest that $PGE_2$ stimulates the differentiation of osteoblasts and generation of osteoclast, and are involved in bone formation, as well as in bone resorption.

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Biocompatibility and Bioactivity of Four Different Root Canal Sealers in Osteoblastic Cell Line MC3T3-El

  • Jun, Nu-Ri;Lee, Sun-Kyung;Lee, Sang-Im
    • Journal of dental hygiene science
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    • v.21 no.4
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    • pp.243-250
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    • 2021
  • Background: Endodontic sealers or their toxic components may become inflamed and lead to delayed wound healing when in direct contact with periapical tissues over an extended period. Moreover, an overfilled sealer can directly interact with adjacent tissues and may cause immediate necrosis or further resorption. Therefore, the treatment outcome conceivably depends on the endodontic sealer's biocompatibility and osteogenic potential. This study aimed to evaluate the cell viability and osteogenic effects of four different sealers in osteoblastic cells. Methods: AH Plus (resin-based sealer), Pulp Canal Sealer EWT (zinc oxide-eugenol sealer), BioRoot RCS (calcium silicate-based sealer), and Well-Root ST (MTA-based calcium silicate sealer) were mixed strictly according to the manufacturer's instructions, and dilutions of sealer extracts (1/2, 1/5 and 1/10) were determined. Cell viability was measured using the water-soluble tetrazolium-8 (WST-8) assay. Differentiation was assessed by alkaline phosphatase (ALP) activity and mineralized nodule formation by Alizarin Red S staining. Results: The cell viability of the extracts derived from the sealers excluding Well-Root ST was concentration dependent, with sealer extracts having the least viability at a 1/2 dilution. At sealer extract dilution of 1/10, the test groups showed the same survival rate as that control group, with the exception of BioRoot RCS. Among all experimental groups, BioRoot RCS showed the highest cell viability after 48 hours. The ALP activity was significantly higher in a concentration-dependent manner. Furthemore, all four materials promoted ALP activity and mineralized nodule formation compared to the control at 1/10 dilutions. Conclusion: This is the first study to highlight the differences in biological activity of these four materials. These results suggest that the composition of root canal sealers appears to alter the form of biocompatibility and osteoblastic differentiation.

Effect of magnesium and calcium phosphate coatings on osteoblastic responses to the titanium surface

  • Park, Ki-Deog;Lee, Bo-Ah;Piao, Xing-Hui;Lee, Kyung-Ku;Park, Sang-Won;Oh, Hee-Kyun;Kim, Young-Joon;Park, Hong-Ju
    • The Journal of Advanced Prosthodontics
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    • v.5 no.4
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    • pp.402-408
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    • 2013
  • PURPOSE. The aim of this study was to evaluate the surface properties and in vitro bioactivity to osteoblasts of magnesium and magnesium-hydroxyapatite coated titanium. MATERIALS AND METHODS. Themagnesium (Mg) and magnesium-hydroxyapatite (Mg-HA) coatings on titanium (Ti) substrates were prepared by radio frequency (RF) and direct current (DC) magnetron sputtering.The samples were divided into non-coated smooth Ti (Ti-S group), Mg coatinggroup (Ti-Mg group), and Mg-HA coating group (Ti-MgHA group).The surface properties were evaluated using scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The surface roughness was evaluated by atomic force microscopy (AFM). Cell adhesion, cell proliferation and alkaline phosphatase (ALP) activity were evaluated using MC3T3-E1 cells. Reverse transcription polymerase chain reaction (RT-PCR) analysis was performed. RESULTS. Cross-sectional SEM images showed that Mg and Mg-HA depositionson titanium substrates were performed successfully. The surface roughness appeared to be similaramong the three groups. Ti-MgHA and Ti-Mg group had improved cellular responses with regard to the proliferation, alkaline phosphatase (ALP) activity, and bone-associated markers, such as bone sialoprotein (BSP) and osteocalcin (OCN) mRNA compared to those of Ti-S group. However, the differences between Ti-Mg group and Ti-MgHA group were not significant, in spite of the tendency of higher proliferation, ALP activity and BSP expression in Ti-MgHA group. CONCLUSION. Mg and Mg-HAcoatings could stimulate the differentiation into osteoblastic MC3T3-E1 cells, potentially contributing to rapid osseointegration.

The biological effects of fibronectin typeIII 7-10 to MC3T3-E1 osteoblast (Fibronectin type III 7-10 이 조골세포에 미치는 영향)

  • Hong, Jeong-Ug;Choi, Sang-Mook;Han, Soo-Boo;Chung, Chong-Pyoung;Rhyu, In-Chul;Lee, Yong-Moo;Ku, Young
    • Journal of Periodontal and Implant Science
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    • v.32 no.1
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    • pp.143-160
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    • 2002
  • 타이태늄은 뛰어난 생체적합성과 적절한 물리적 성질을 바탕으로 치과 및 정형외과 영역의 매식재로 널리사용되어져 왔으며, 골과 매식재 사이의 골 융합 정도를 증가시킬 목적으로 물리, 화학적인 방법을 이용한 타이태늄의 표면처리에 관한 많은 연구들이 진행되어 왔다. 최근에는 부착단백질 또는 성장인자를 이용한 생체재료의 표면개질을 통하여 조직적합성 및 치유 능의 개선을 위한 시도들이 있어왔다. Fibronectin(FN)은 주요 세포외기질중의 하나로 생체 내 널리 분포하여 세포의 부착, 이동 및 증식에 관여하는 거대 당단백으로, RGD및 PHSRN 펩타이드 서열이 세포의 인테그린과 결합하여 세포의 활성을 조절하는 것으로 알려져 있다. 이 연구에서는 FN으로 처리된 타이태늄이 조골세포의 부착, 증식 및 분화에 미치는 영향과 이에 따른 석회화 정도에 미치는 영향을 관찰하여 부착분자를 이용한 타이태늄 표면개질의 효과를 규명하고자 하였다. 상업용 순수 타이태늄을 gold thiol법을 이용하여 표면처리 후, 혈장 FN(plasma FN, pFN)과 유전자재조합법을 이용하여 얻은 FN조각(FN type III 7-10, FNIII 7-10)을 피복한 시편을 실험군으로, 아무런 처리를 하지 않은것(smooth surface, SS)과 산 부식(Sandblasted and acid etched, SLA)처리된것을 대조군으로 이용하였다. 배양된 조골세포주(MC3T3-E1)를 사용하여 타이태늄 표면 처리에 따른 세포의 증식, 형태변화, 알칼리성 인산분해효소(ALPase) 생산 및 세포면역형광법을 이용한 분화정도를 시간 경과에 따라 관찰하였다. 조골세포증식의 경우 FNIII 7-10 처리군에서 pFN 처리군 및 대조군에 비해 시간경과에 따라 유의성있는 세포수의 증식이 관찰되었으며(p<0.05), ALPase 생성의 경우에도 FNIII 7-10 처리 군에서 아무 처리도 하지 않은 군에 비해 유의성 있게 높은 효소의 생성이 관찰되었다(p<0.05). 주사전자현미경을 이용한 세포의 형태관찰결과 아무 처리도 하지 않은 군에서는 마름모형태를 나타내었으며, 산 부식 처리된 군에서는 세포가 가시모양의 형태를 보인 반면 FN으로 처리된 두 군에서는 세포의 부착 및 펴짐이 매우 발달되어 있는 모습이 관찰되었다. 세포의 분화정도를 관찰하기 위하여 국소부착키나제(focal adhesion kinase, FAK), 및 actin stress fiber의 분포양상을 세포면역형광법을 이용하여 관찰한 결과 FN으로 표면처리된 두 군에서 아무런 처리도 하지않은 군 및 산 부식처리 한 군에 비해 프라크의 발현이 높게 나타났으며 잘 발달된 actin stress fiber의 소견을 나타내었다. 이 실험의 결과들은 gold thiol 법을 이용한 표면처리 후 FN부착을 통한 타이태늄의 표면개질이 조골세포의 부착, 증식 및 분화에 중요한 역할을 담당하여 석회화 정도를 촉진시키는 것을 보여주었으며, 이런 결과들은 더 짧은 FN조각을 이용한 다른 생체재료의 표면개질에 폭 넓게 응용할 수 있으리라 생각된다.

Surface characteristics and bioactivity of an anodized titanium surface

  • Kim, Kyul;Lee, Bo-Ah;Piao, Xing-Hui;Chung, Hyun-Ju;Kim, Young-Joon
    • Journal of Periodontal and Implant Science
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    • v.43 no.4
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    • pp.198-205
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    • 2013
  • Purpose: The aim of this study was to evaluate the surface properties and biological response of an anodized titanium surface by cell proliferation and alkaline phosphatase activity analysis. Methods: Commercial pure titanium (Ti) disks were prepared. The samples were divided into an untreated machined Ti group and anodized Ti group. The anodization of cp-Ti was formed using a constant voltage of 270 V for 60 seconds. The surface properties were evaluated using scanning electron microscopy, X-ray photoelectron spectroscopy, and an image analyzing microscope. The surface roughness was evaluated by atomic force microscopy and a profilometer. The contact angle and surface energy were analyzed. Cell adhesion, cell proliferation, and alkaline phosphatase activity were evaluated using mouse $MC_3T_3-E_1$ cells. Results: The anodized Ti group had a more porous and thicker layer on its surface. The surface roughness of the two groups measured by the profilometer showed no significant difference (P>0.001). The anodized Ti dioxide ($TiO_2$) surface exhibited better corrosion resistance and showed a significantly lower contact angle than the machined Ti surface (P>0.001). Although there was no significant difference in the cell viability between the two groups (P>0.001), the anodized $TiO_2$ surface showed significantly enhanced alkaline phosphatase activity (P<0.001). Conclusions: These results suggest that the surface modification of Ti by anodic oxidation improved the osteogenic response of the osteoblast cells.

Preparation of Sulfobetaine Chitosan, Silk Blended Films, and Their Properties (설포베타인 키토산의 실크 블렌드 필름의 제조 및 그들의 성질)

  • Koo, Ja-Sung;Cha, Jae-Ryung;Oh, Se-Heang;Gong, Myoung-Seon
    • Polymer(Korea)
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    • v.38 no.1
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    • pp.54-61
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    • 2014
  • Water-soluble sulfobetaine chitosan (SCs) was prepared for a blending film with Bombyx mori silk fibroin (SF) by reacting chitosan with 1,3-propanesultone. A series of SF/SCs blended films were successfully prepared by mixing aqueous solutions of B. mori SF and SCs. The SF/SCs blended films were examined through spectroscopic and thermal analysis to determine the morphological changes of SF in the SCs. The effects of the SF/SCs blend ratios on physical and mechanical properties were investigated to discover the feasibility of using these films as biomedical materials such as artificial skin and wound dressing. X-ray analysis showed good compatibility between the two biopolymers. The in vitro degradation behavior of the SF/SCs blended films was systematically investigated for up to 8 weeks in phosphate buffered saline solution at $37^{\circ}C$ and showed a mass loss of 46.4% after 8 weeks. All films showed no cytotoxicity by MC3T3-E1 assay. After 3 days of culture, the relative cell number on all the SF/SCs films was slightly lower than that of an optimized tissue culture plastic.