Biomedical subjects

Chandra P Sharma

Publications and source records attributed to Chandra P Sharma.

11 recordsLinked to original sources

PubMed2006

Effect of calcium, zinc and magnesium on the attachment and spreading of osteoblast like cells onto ceramic matrices.

Calcium phosphate ceramic has been widely used as bone substitute materials. Neumerours approaches have been investigated to develop tissue-engineered scaffold from hydroxyapatite because of its advantages like osteoconduction. We have developed porous ceramic matrices from nanoparticles of calcium phosphate containing zinc and magnesium. Mimicking the grain size of natural bone enhances the bone forming function of cells. Osteoblast-like MG63 cells were cultured on to these porous ceramic matrices. Cell adhesion and spreading onto these matrices were studied for 24 h and 5 days in vitro. It was observed that on calcium phosphate matrix, containing a combination of zinc and magnesium, the osteoblast adhesion and spreading was significant on 5th day. This appeared to be comparable to the hydroxyapatite control. This makes it a promising candidate as a bone tissue-engineering scaffold.

Bone Substitutes↗

PubMed2006

Cyclodextrin-insulin complex encapsulated polymethacrylic acid based nanoparticles for oral insulin delivery.

Present investigation was aimed at developing an oral insulin delivery system based on hydroxypropyl beta cyclodextrin-insulin (HPbetaCD-I) complex encapsulated polymethacrylic acid-chitosan-polyether (polyethylene glycol-polypropylene glycol copolymer) (PMCP) nanoparticles. Nanoparticles were prepared by the free radical polymerization of methacrylic acid in presence of chitosan and polyether in a solvent/surfactant free medium. Dynamic light scattering (DLS) experiment was conducted with particles dispersed in phosphate buffer (pH 7.4) and size distribution curve was observed in the range of 500-800 nm. HPbetaCD was used to prepare non-covalent inclusion complex with insulin and complex was analyzed by Fourier transform infrared (FTIR) and fluorescence spectroscopic studies. HPbetaCD complexed insulin was encapsulated into PMCP nanoparticles by diffusion filling method and their in vitro release profile was evaluated at acidic/alkaline pH. PMCP nanoparticles displayed good insulin encapsulation efficiency and release profile was largely dependent on the pH of the medium. Enzyme linked immunosorbent assay (ELISA) study demonstrated that insulin encapsulated inside the particles was biologically active. Trypsin inhibitory effect of PMCP nanoparticles was evaluated using N-alpha-benzoyl-L-arginine ethyl ester (BAEE) and casein as substrates. Mucoadhesive studies of PMCP nanoparticles were conducted using freshly excised rat intestinal mucosa and the particles were found fairly adhesive. From the preliminary studies, cyclodextrin complexed insulin encapsulated mucoadhesive nanoparticles appear to be a good candidate for oral insulin delivery.

2-Hydroxypropyl-beta-cyclodextrin↗

PubMed2006

Cell mimetic lateral stabilization of outer cell mimetic bilayer on polymer surfaces by peptide bonding and their blood compatibility.

The biological lipid bilayer membranes are stabilized laterally with the help of integral proteins. We have simulated this with an optimized ternary phospholipid/glycolipid/cholesterol system, and stabilized laterally on functionalized poly methyl methacrylate (PMMA) surfaces, using albumin, heparin, and polyethylene glycol as anchors. We have earlier demonstrated the differences due to orientation and packing of the ternary phospholipid monolayers in relation to blood compatibility (Kaladhar and Sharma, Langmuir 2004;20:11115-11122). The structure of albumin is changed here to expose its interior hydrophobic core by treating with organic solvent. The interaction between the hydrophobic core of the albumin molecule and the hydrophobic core of the lipid molecules is confirmed by incorporating the molecule into bilayer membranes. The secondary structure of the membrane incorporated albumin is studied by CD spectral analysis. The structure of the altered albumin molecule contains more beta-sheet as compared to the native albumin. This conformation is also retained in membranes. The partitioning of the different anchors based on its polarity and ionic interactions in the monolayer is studied from the pressure-area (pi-A) isotherm of the lipid monolayers at the air/water interface using Langmuir-Blodgett (LB) trough facility. Such two monolayers are deposited onto the functionalized PMMA surface using LB trough and crosslinked by carbodiimide chemistry. The structure of the deposited bilayer is studied by depth analysis using contact mode AFM in dry conditions. The stabilized bilayer shows stability up to 1 month by contact angle studies. Preliminary blood compatibility studies reveal that the calcification, protein adsorption, as well as blood-cell adhesion is significantly reduced after the surface modification. The reduced adsorption of ions, proteins, and cells to the modified surfaces may be due to the fluidity of the microenvironment along with the contribution of the mobile PEG groups at the surface and the phosphorylcholine groups of the phospholipids. The stability of the anchored bilayer under low shear stress conditions promises that the laterally stabilized supported bilayer system can be used for low shear applications like small diameter vascular graft and modification of biosensors, and so forth.

Animals↗

PubMed2006

Cell mimetic monolayer supported chitosan-haemocompatibility studies.

Chitosan is a natural polymer, widely explored for biomedical and tissue engineering applications. However the thrombogenic nature limits their application in blood contacting devices and implants. Here, we have attempted to understand the haemocompatibility of chitosan by immobilizing a monolayer of cell mimetic lipid compositions. The phosphatidylcholine/cholesterol/galactocerebroside lipid composition (PC/Chol/GalC, 1:0.35:0.125) was deposited onto the chitosan films. Characterization of the modified surface was done by sessile drop contact angle measurement. The contact angle of the chitosan film reduced from 80.65 +/- 1.4 to 23.5 +/- 1.9 after the surface modification. Swelling nature of chitosan seemed to influence the orientation and packing of the lipid monolayer. In vitro calcification studies with metastable salt solution indicated increased calcification on the modified surface. This may be due to formation of nuclei for calcification on the expanding monolayer. The preliminary haemocompatibility studies with washed platelets, leukocytes and erythrocytes showed overall reduction in blood cell adhesion to the modified surfaces. Scanning electron microscopy was used for morphological characterization of platelet adhesion and activation on the surfaces. On the bare chitosan surface, fully spread platelets with extending pseudopodia indicated platelet activation. The smooth surface of the modified film did not activate platelets. These studies showed that, though the lipid monolayer on chitosan film is able to reduce the over all blood cell adhesion and platelet activation it is prone to calcification.

Biocompatible Materials↗

PubMed2006

Novel pH responsive polymethacrylic acid-chitosan-polyethylene glycol nanoparticles for oral peptide delivery.

In present study, novel pH sensitive polymethacrylic acid-chitosan-polyethylene glycol (PCP) nanoparticles were prepared under mild aqueous conditions via polyelectrolyte complexation. Free radical polymerization of methacrylic acid (MAA) was carried out in presence of chitosan (CS) and polyethylene glycol (PEG) using a water-soluble initiator and particles were obtained spontaneously during polymerization without using organic solvents or surfactants/steric stabilizers. Dried particles were analyzed by scanning electron microscopy (SEM) and particles dispersed in phosphate buffer (pH 7.0) were visualized under transmission electron microscope (TEM). SEM studies indicated that PCP particles have an aggregated and irregular morphology, however, TEM revealed that these aggregated particles were composed of smaller fragments with size less than 1 micron. Insulin and bovine serum albumin (BSA) as model proteins were incorporated into the nanoparticles by diffusion filling method and their in vitro release characteristics were evaluated at pH 1.2 and 7.4. PCP nanoparticles exhibited good protein encapsulation efficiency and pH responsive release profile was observed under in vitro conditions. Trypsin inhibitory effect of these PCP nanoparticles was studied using casein substrate and these particles displayed lesser inhibitory effect than reference polymer carbopol. Preliminary investigation suggests that these particles can serve as good candidate for oral peptide delivery.

Administration, Oral↗

PubMed2004

Supported cell mimetic monolayers and their interaction with blood.

Surface modification using supported monolayers of phosphorylcholine containing phospholipids has been an accepted strategy for developing blood-contacting materials. We present a detailed study of the blood compatibility of the supported monolayers of phospholipid, glycolipid, and cholesterol (Chol) binary and ternary lipid combinations using in vitro techniques. The packing and orientation of these monolayers have been correlated with the blood compatibility. We have used phosphatidylcholine (PTC) for phospholipid, galactocerebroside (Gal) for glycolipid, and Chol based on the headgroup structure to represent the major lipid components of the endothelial luminal cell membrane. The interfacial behavior of various combinations of PTC, Gal, and Chol monolayers have been studied at the air/water interface and deposited on hydrophobic polycarbonate (PC) polymer substrates with the help of the Langmuir-Blodgett trough. The packing and orientation of the supported monolayers have been varied by means of changing the lipid composition rather than the deposition parameters. This approach seems to be more similar to the in vivo conditions. The different supported monolayer surfaces prepared accordingly are (1) a closely packed ordered hydrophobic surface, PC modified with the combination PTC/Chol/Gal (1:0.35:0.125), (2) a loosely packed ordered hydrophobic surface, PC modified with the combination PTC/Chol (1:0.35), and (3) a closely packed ordered hydrophilic surface, PC modified with the combination PTC/Chol (1:0.7). An optimized modified surface (PTC/Chol/Gal, 1:0.35:0.125) has been identified on the basis of the maximum transfer ratio from the air/water interface and characterized by using atomic force microscopy. The concentration of Chol has been found to be an important parameter, which influences the transfer ratio. The Gal improves the monolayer integrity under a reduced Chol concentration. The blood compatibility of these supported monolayers was studied by protein adsorption, blood cell adhesion, and calcification. The tightly packed ordered hydrophobic surface (PTC/Chol/Gal, 1:0.35:0.125), has been found to be more blood compatible because of reduced blood cell adhesion and calcification. This surface also promotes albumin adsorption and may be the reason for the reduced platelet activation, while in the case of the loosely packed ordered hydrophobic surface (PTC/Chol, 1:0.35) the protein adsorption also has been reduced along with the blood cell adhesion and calcification. When the ordered hydrophilic surface (PTC/Chol, 1: 0.7) of the monolayer has been exposed, the blood cell adhesions as well as the overall protein adsorption were significantly reduced. However, the packing of the phosphorylcholine moieties of the polar headgroup has been affecting the calcification on the surface. We have observed an increase in calcification to the surface modified with the loosely packed polar headgroup, from a relative study on chitosan and chitosan modified with the monolayer of PTC. These findings are helpful for the surface modifications for blood-contacting materials using this strategy.

Air↗

PubMed2004

Poly methacrylic acid-alginate semi-IPN microparticles for oral delivery of insulin: a preliminary investigation.

Microparticles of Poly methacrylic acid (P1) and novel semi-interpenetrating network composed of Poly methacrylic acid-alginate (P2) were prepared and their application in oral insulin delivery was evaluated. The microparticles were characterized by scanning electron microscopy (SEM) for morphological studies. Insulin loading onto the microparticles was performed by the diffusion filling method and insulin encapsulated microparticles were subjected to in vitro release study in buffer solution of pH 1.2 and 7.4. The release kinetics at pH 7.4 exhibited sustained release of insulin for more than 5 h in case of PMAA microparticles whereas burst release of insulin (90% of total insulin loaded) within 1 h of study was observed in the case of PMAA-alginate microparticles. At pH 1.2, around 30% of insulin loaded was released from both microparticles within 2 h of study.

Administration, Oral↗

PubMed2003

Oral insulin--a perspective.

Diabetes mellitus is generally controlled quite well with the administration of oral medications or by the use of insulin injections. The current practice is the use of one or more doses, intermediate or long acting insulin per day. Oral insulin is a promising yet experimental method providing tight glycemic control for patients with diabetes. A biologically adhesive delivery systems offer important advantage over conventional drug delivery systems. The engineered polymer microspheres made of erodable polymer display strong adhesive interactions with gastrointestinal mucus and cellular lining can traverse both the mucosal epithelium and the follicle associated epithelium covering the lymphoid tissue of Peyer's patches. Alginate, a natural polymer recovered from seaweed is being developed as a nanoparticle for the delivery of insulin without being destroyed in the stomach. Alginate is in fact finding application in biotechnology industry as thickening agent, a gelling agent and a colloid stabilizer. Alginate has in addition, several other properties that have enabled it to be used as a matrix for entrapment and for the delivery of a variety of proteins such as insulin and cells. These properties include: a relatively inert aqueous environment within the matrix; a mild room temperature encapsulation process free of organic solvents; a high gel porosity which allows for high diffusion rates of macromolecules; the ability to control this porosity with simple coating procedures and dissolution and biodegradation of the system under normal physiological conditions.

Administration, Oral↗

PubMed2003

Ceramic drug delivery: a perspective.

Different ceramic substances are offered in the market as bone substitute materials. These include monophasic calcium phosphate ceramics of tricalciumphosphate (TCP) or hydroxyapatite (HA), biphasic calcium phosphate ceramics and multiphasic bio-glasses synthetic calcium phosphate cements. Ceramics with appropriate three-dimensional geometry are able to bind and concentrate bone morphogenetic proteins in circulation and may become osteoinductive (capable of osteogenesis) and can be effective carriers of bioactive peptide or bone cell seeds and are therefore potentially useful in tissue engineering and drug delivery. An attempt has been made to review various drug delivery applications of ceramics.

Biocompatible Materials↗

PubMed2002

Delivery of insulin from hydroxyapatite ceramic microspheres: preliminary in vivo studies.

The major limiting factor in utilizing protein drugs for sustained delivery is the lack of suitable delivery systems. Ceramic hydroxyapatite microspheres are biocompatible and utilized for the purification of proteins. As a preliminary study, we have investigated the possibility of using hydroxyapatite ceramic microspheres loaded with insulin as an implantable delivery system in rats. With this limited test, it was shown that the loaded insulin was active and able to suppress the blood glucose level in normal rats.

Animals↗

PubMed2002

Stimuli sensitive polymethacrylic acid microparticles (PMAA)--oral insulin delivery.

This study investigated polymethacrylic acid (PMAA) microparticles for controlled release of Insulin in oral administration. The microparticles were characterised by scanning electron microscopy (SEM) for morphological studies. The swelling behaviour and drug release profile in various pH media were studied. The % swelling of gels was found to be inversely related to the amount of crosslinker added. Inclusion complex of betaCD and Insulin was studied using polyacrylamide gel electrophoresis (PAGE). Optimum complexation was obtained in the ratio 100 mg betaCD: 200 IU Insulin. The release pattern of Insulin from Insulin-betaCD complex encapsulated PMAA microparticles showed release of Insulin for more than seven hours.

Administration, Oral↗