Interaction Between Nanohydroxyapatite And Biomacromolecule And Erythrocyte In Blood

As a high-quailty medical biomaterials,hydroxyapatite have been applicated on artificial bone and implant dentes. Nano-hydroxyapatite became investigative warm spots in biomedicine domain such as artificial bone, medicine carrier and antineoplastic activity for its nanometer size effectiveness and fine biocompatibility.But the safe study of Nano- hydroxyapatite in blood is the key link to come true clinical application which is reported lesser currently.This paper have approached interaction between Nano- hydroxyapatite and some of important biomolecules in blood and its influence on morphous and function of red blood corpuscle(RBC). Ultimum,We hope to provide some foundational base on reshaping of material surface for raising haemo- compatibility.In this study, different nano-hydroxyapatite particles,HAP₁(25-60nm), HAP₄(the additives is heparin, 15-50nm), HAP₅(the additives is bovine serum albumin BSA, 20-80nm) were prepared by homogeneous precipitation and used heavy-gauge hydroxyapatite as comparison,we determined amount of heparin, sialic acid ,BSA adsorbed on HAP₁,HAP₂,HAP₃ by Crystal Violet assay, Bialsche method,Bradford method respectively and analyzed the binding mechanism by infrared spectrum;After taking HAP₁,HAP₂,HAP₄,HAP₅ and RBC to co-culture in vitro,we studied RBC hemolysis test and detected RBC hematolysis rate by erythrocyte osmotic fragility test;observing the changes of morphous and locomotion of cell after coacting HAP₁,HAP₂,HAP₄,HAP₅ and RBC by light microscope and inverted phase contrast microscope;observing HAP₁,HAP₂,HAP₄,HAP₅ effecting on Ultrastructure of RBC.Eventually,we have the conclusions below: 1 .There are different extent absorption characteristics between HAP₁,HAP₂,HAP₃ and heparin, sialic acid ,BSA while the adsorption amount of HAP₁ higher away than HAP₂ and HAP₃ because of distinct nanometer potency. Furthermore, HAP₁ adsorbed heparin by [Ca²+] binding [-OSO3-],[COOH] binding [OH] via hydrogen bond and molecular structure curl and overlapping of heparin;HAP₁ adsorbed sialic acid chief by [OH^-] combining [NH] and [Ca²+] combining [COOH^];There have almost no difference on adsorption amount of sialic acid adsorbing onto HAP₁ between heparinized and unheparinized HAP₁ The adsorptional site of HAP₁ and BSA are primary at acid amide I, acid amideII, [COO"] group of BSA and [Ca2+] group, [OH] group, [PO43'] group of hydroxyapatite.2.HAPi,HAP2,HAp4,HAP5 have not caused hematolysis after they co-cultured with RBC,but HAPi and HAP4can lead to erythrocyte aggregation while HAP5 can keep RBC floating.3.From ultrastructure of RBC which co-cultured with HAP|,HAP4,HAP5,we have observed that HAPi and HAP4 can induce RBC membrane to deform and decrease stability of RBC membrane, so the particles can enter intramembrane, as a result, intracellular hydroxyapatite nanoparticles may heighten the concentration of Ca2+ and effect structure of membrane protein, membrane lipid and hemoglobin, subsequently,the deformability of RBC decreased all the better and leaded to erythrocyte aggregation. However,HAP.s uniformly dispersed around cell and didn't adhere to it,also, cytomorphosis cannot be found.In short, HAPi,HAP2,HAP4,HAP5 didn't lead to plasmatorrhexis after they interacted with RBC, further, HAP5 with BSA ostensible modifier is ideal haemo-compatibility materials because it can maintain RBC's normal morphous and floaty nature.