Modification of chitosan membrane with poly(vinyl alcohol) and biocompatibility evaluation
Highlights
► Chitosan had strong interactions with poly(vinyl alcohol) by forming hydrogen bond. ► Adding PVA into chitosan membrane could improve its flexibility and wettability. ► MTS cytotoxicity essay indicated CS/PVA membranes were biocompatible. ► Cells attachment studies showed CS/PVA membranes were biocompatible.
Introduction
Tissue engineering has been regarded as an ultimate medical treatment for reconstruction of defective tissues in biomedical engineering fields in recent years [1]. Guided tissue regeneration (GTR) is a comparatively simple tissue engineering treatment compared with other treatments such as in vitro tissue reconstruction, which reconstructs new tissues by using a barrier membrane to prevent other tissues from invading the defected areas [2], [3]. Recently GTR and relative biomaterials used in GTR treatment have being paid an increasing attention by a great number of researchers [4], [5], [6], [7], [8]. The materials used in GTR applications mainly include nondegradable and degradable materials. Though expanded polytetrafluoroethylene membrane (e-PTFE) as an exemplary nondegradable membrane has achieved excellent clinical results, it needs second surgery procedure to remove the membrane after the regeneration of new bone, which increases both the risk of surgery and the financial burden of patients. In order to solve the drawbacks of nondegradable membranes, people turn to use degradable membranes such as poly lactic acid (PLA), polyglycolic acid (PLG), its copolymer (PLGA) and Collagen membranes. However, these kinds of membranes have their drawbacks. Acid products will be produced during the degradation and resorption process of PLA, PLG. The accumulation of acid products will significantly decrease the pH value, which will cause inflammatory response [9], [10]. Collagen membrane usually has weak strength and is difficult to manipulate. Besides, the resorption rate is hard to match with normal tissue-healing process. As a result, GTR membranes with good biocompatibility, proper mechanical properties and suitable degradation rate are needed to be produced.
In order to fulfill the demands of GTR technology, one good way is utilizing the composites of synthetic and natural biodegradable polymers, which combines the good mechanical properties and process ability of synthetic polymers with the specific tissue and cell compatibility of biopolymers, thus receiving increasing interest owning to their potential applications in biomedical field [11].
The aim of this work is to find one kind of more suitable biomaterials prepared from CS and PVA composite to satisfy the requirements of GTR technology. CS [poly(1,4),-β-d-glucopyranosamine] is the second abundant natural polysaccharide, which has many useful properties: nontoxicity, biocompatibility, biodegradability, bioactivity, antibacterial, excellent film forming capacity [12], [13], [14]. Thus CS is widely used in a number of biomedical applications including drug delivery systems [15], [16], tissue engineering [17], [18], wound dressing [19]. However the major imperfections of chitosan such as poor workability and high brittleness [20] cause it impractical in real biomedical applications, so the selection of some helpful additives with good compatibility and mechanical properties is necessary. An effective method to overcome chitosan's drawbacks is to blend it with a synthesized polymer to develop a composite. As a synthesized polymer, PVA possesses many excellent properties: highly hydrophilic properties, excellent mechanical properties, nontoxicity, water-solubility, biocompatibility, and biodegradability. As a result, it has been widely used in biomedical field [21]. To combine the excellent properties of PVA with those of CS, CS/PVA membranes have been developed. Although a few researchers have studied CS/PVA blend films by solution casting method, all of these studies were used in other fields such as food packaging [22] instead of GTR. In order to study the possibility of use of CS/PVA membranes as GTR membrane, a thorough and systematic study of CS/PVA membranes is needed. So in this work, the chemical structure, crystalline, thermal properties, mechanical properties and wettability of CS/PVA membranes were studied by FTIR, XRD, DSC, tensile testing and contact angle instrument, respectively. The biocompatibility of CS/PVA membranes was also evaluated by MTS cytotoxicity assay and cell attachment test.
Section snippets
Materials
CS (biomedical grade, Mη = 5.63 × 105, D.D = 91%) was purchased from Qingdao Haihui Bioengineering Co., Ltd., China. Acetic acid (CP) was purchased from Yixing Niujia Chemical Reagent plant, China. NaOH (AR) was obtained from Hangzhou Xiaoshan Chemical Reagent Corporation, China. PVA-124 (AR, Mn = 1.06 × 105) was obtained from Sinopharm Chemical Reagent Co., Ltd., China. Trypsin–EDTA, Dulbecco's modification of Eagle's medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco, Invitrogen
FT-IR spectra
To study the compatibility of CS/PVA membranes and understand the interactions between CS molecules and PVA molecules, FT-IR measurements were taken (see Fig. 1).
The spectra of CS membrane and CS/PVA membranes with different weight ratio showed –O–H and –N–H stretching at about 3427 cm−1, –C–H stretching at about 2921 cm−1 and –C–O stretching at about 1089 cm−1. For pure CS membrane, a peak at 1596 cm−1 appeared which was attributed to the bending vibration of –NH2. While the peak at about 1651 cm−1
Conclusions
In this study, CS membrane and CS/PVA membranes with different weight ratio were prepared by solution casting method. CS/PVA membranes’ chemical, crystalline and thermal properties were studied by FT-IR, XRD and DSC. CS and PVA had strong interactions with each other by forming hydrogen bonds between them. Adding PVA into CS membrane could greatly modify its flexibility and wettability. MTS cytotoxicity assay indicated that CS/PVA membranes had good biocompability. Cell attachment studies
Acknowledgements
This work was funded by National Science Foundation of China (grant no. 50773070), The Key Basic Research Development Plan of China (grant no. 2009CB930104), Grand Science and Technology Special Project of Zhejiang Province (grant no. 2008C11087). The authors thank Associate Professor Mao Peng and Honglei Guo for their kind support of tensile testing in their laboratory.
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