Biocom pa t ibilit y of Ch it osa n / Colla ge n / PVA N a n ocom posit e
substance including glycosam inoglycans, proteoglycans, and glycoproteins, while t he inorganic phase is com posed of calcium phosphate such as hydroxyapat it e and - t ricalcium phosphate ( Zhao et al., 2002) . I n this works, bioceram ic of bovine bone, chit osan, collagen and poly( vinyl alcohol) were chosen to prepare t he organic- inorganic com posites.Bovine bone as biological wast e is very m any found in I ndonesia. I t s ut ilizat ion as biom aterial in bone t issue engineering is st ill less. Bovine bone is m ore widely used as supplem entary food for livest ock. st ruct ure is sim ilar t o glycosam ino glycans and hyaluronic acid found in cart ilage ( Risbud et al., 2005).
Collagen is a fibrous protein hollow and t here are lot s of j oint t issues, skin, m uscles, and tendons. This nat ural fibrous connect s and support s ot her bodily t issues, such as bone and tendon (Hossein et al., 2007) . I n t he field of derm at ology collagen is used as a growth enhancer net work. Polyvinyl alcohol is a sem i- cryt alline synthet ic polym er t hat wat er-soluble, non- toxic, non-carcinogenic, biocom pat ible and elast ic physical propert ies. I n addit ion PVA also has a st rong adhesive power so it is good as a com posit e m at rix because it can im prove t he com pact ness and m echanical propert ies of t he com posit e. These charact erist ics m ake t his polym er part icularly suitable for biom edical applicat ions.
I n t his st udy, bioceram ic was prepared by calcining bovine bones and crushed by ball m illing and ult rasonicat ion in aquabidest . Chit osan- collagen-poly( vinyl alcohol) / bioceram ic com posit es were prepared by m echanical m ixing and dried using freeze dryer. The m orphology and physicalchem ical propert ies of t he com posites such as t he porosity, degradat ion and com pressive st rengt h were invest igated. I n addit ion, also t he biocom pat ibilit y of t he com posites was prelim inary evaluated by in vivo im plant at ion int o t he fem oral m uscle of whit e m ouse.
M a t e r ia ls a nd M e t h ods
Bioceram ic was t ypically prepared by calcining bovine bone as follows. First ly, bovine leg bones were wit h hem at oxylin and eosin (HE) reagen and observed under an opt ical m icroscope.
Resu lts a nd D iscussion
Preparat ion of Bioceram ic and Chitosan-Collagen-Poly( Vinyl Alcohol) / Bioceram ic Com posites
I n t he bioceram ic preparat ion, 121alcinat ions process on bovine bone aim s t o obt ain pure calcium -before being processed. The com posites are obtained seem hom ogeneous and also have pores.
Charact erizat ions
com posit e possesses cont rolled biodegradat ion, which is suit able for a biom at erial for bone graft subst it ut e.
Figure 1. Biodegradat ion of chit osan- collagen-poly ( vinyl alcohol) / bioceram ic com posites.
Mechanical Propert ies
Mechanical propert ies, especially com pressive st rength, are im port ant and usually im perat ive crit erion in select ing t he bone graft subst it ut e m aterials ( Yin et al., 2003; Panzavolta et al., 2009) . Hence, t he com pressive st rength of t hese m aterials were shown in Figure 2. From the records, it could be found t hat t he com pressive st rengt h of t he com posites had a relat ion t o t he bioceram ic content in com posit es which t he m echanical propert ies increased by rise of m ass of bioceram ics from 4 unt il 6 g in t he com posites. However, t he m echanical propert ies decreased by adding t he bioceram ics m ore t han 6 g or when t he m ass of the bioceram ics exceeded 7 g. it m eans t hat presence of sm all am ount of t he bioceram ics could im prove the m echanical propert ies of t he com posites but t he large am ount of t hose could reduce t he degree of t he m echanical propert ies. Zhou et al.. ( 2012) reported t hat com pressive st rengt h of gelat in/ CM-chit osan/- TCP declined wit h t he addit ion of fract ion of - TCP m ore t han 5% . I n t his study, t he com posit e wit h 6 g m ass of bioceram ics is t he com posit e t hat has t he com pressive st rengt h in range of t he degree of cancellous’s com pressive st rengt h which is 2- 12 MPa (Ficai, et al., 2011) . I t was pot ent ial used as bone graft subst it ute m at erials.
Figure 2. St ress- St rain curve of chit osan- collagen-poly ( vinyl alcohol) / bioceram ic com posites.
Fourier Transform I nfrared Spect roscopy
FTI R spect ra were recorde t o a bet ter underst anding of t he int eract ions bet ween t he com ponents in t he com posit es. The FTI R spect ra of chit osan, collagen, poly( vinyl alcohol) , bioceram ic, and t he com posit es before and after degraded were shown in Figure 3.
C4
C5
C6
4000 3500 3000 2500 2000 1500 1000 500 seem s t he m agnit ude of t hese bands becom es weaker. The absorpt ion bands of chit osan and collagen such as am ide A, am ide B, am ide I , am ide I I , am ide I I I and COO-, and also v1 m ode of PO43- and v5 m ode of PO43- are absent in t he com posit es ( Figure 3e) . t hese spect ral changes and absent s indicate t here have been interact ions bet ween t he com ponents.
The com posit e after degraded for 7 days has sim ilar FTI R spect ra ( Figure 3f) wit h FTI R spect ra of the
Figure 4. SEM im ages of (a) bioceram ic and (b) chit osan-collagen- poly( vinyl alcohol) / bioceram ic com posit e with 6 g bioceram ic cont ent .
Biocom pat ibilit y of chit osan-collagen-poly( vinyl alcohol) / bioceram ic com posite
Biocom pat ibilit y t est s of t he nanocom posit es as bone graft were carried out using live m ice wounded t issue after t reatm ent for 7 and 14 days.
( 1) ( 2)
Figure 5. Mice t issue wound healing on day- 7 ( 1) and day- 14 ( 2) aft er im plantat ion of t he nanocom posit e as bone graft
Con clusions
Mineral of calcium apet ite has been successfully isolat ed from bovine bone waste by calcinat ion at 830oC and after ballm illing and charact erizat ion using part icle size analyser ( PSA) it was found t hat t he part icle size was ranging 100 - 8000 nm . Nanocom posites of t he calcium apet ite- filled chit osan-collagen showed good m echanicl propert ies ( com pression st rengt h: 2.01 MPa and m ore im portant ly were biocom pat ible wit h live m ice t issue without indict ion of t oxicit y
.
Re fe r e n ce s
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