Assessment of the effect of biodegradable implants based on polylactide on the organism animals by analysis of the dynamics of biochemical indicators of the blood at different periods of research

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Assessment of the effect of biodegradable implants based on polylactide on the organism animals by analysis of the dynamics of biochemical indicators of the blood at different periods of research

osteosynthesis calcium phosphate

Pavlov Oleksiy Dmytrovych

Аssistant of the Traumatology and Orthopedics, Physical and Rehabilitation Medicine Department

Kharkiv National Medical University, Ukraine

Pavlova Olena Oleksiivna

Doctor of Medicine, Professor, Professor of the D.O. Alpern General and Clinical Рathophysiology Department

Kharkiv National Medical University, Ukraine

Summary

In experimental conditions, at different times (14th, 30th, 90th, 180th, and 360th day), with the help of modern methods, the general effect on the body of rats of implants for osteosynthesis based on polylactide and calcium phosphate was investigated ceramics In comparison with the control, it was established that: the composite material, in general, does not affect the somatic status of the animals in all terms of the experiment. The content and activity of biochemical markers of the regenerative-inflammatory process (glycoproteins, chondroitin sulfates, alkaline phosphatase) in the blood serum of rats on the 30th day after implantation (composite based on polylactide) indicate an easier course of the process of restoring the injured bone, and indicators of the functional state of the liver ( ALT, AST, GHTP, bilirubin) and kidneys (creatinine, urea) after implantation in both studied groups of rats, indicates the absence of nephro- and hepatotoxicity of the implants in all periods of observation.

Key words: experiment, rats, polylactide, calcium-phosphate ceramics, implants, femur, biochemical methods,

The percentage of injuries to the locomotor system ranks second among the causes of injuries and third among diseases that lead to disability in the adult population [1 ]. The clinical efficiency of medical institutions in this direction requires and economically justifies the introduction of new technological developments, among which the manufacture of implants for osteosynthesis occupies a special place. The properties of materials for the implementation of such a task must meet certain requirements: not to enter into undesirable chemical reactions with tissues and interstitial fluid, to be resistant to corrosion, strong and wear-resistant, not to cause reactions from the immune system, promote the consolidation of fragments and have a positive effect on osteogenesis [2] contribute to a significant improvement in the quality of life of patients. Therefore, there is a need to study the effect of improved biodegradable implants on the functions of the animal body at different times after implantation, which determines the study's relevance.

The purpose of the study is to evaluate the dynamics of the biochemical indicators of the blood of rats after the introduction of a new bioediting composite material based on polylactide into their femur to determine the consequences of its influence on the organism.

Materials and methods. Implantation of a new material based on polylactide and calcium-phosphate ceramics in the femoral compartment was carried out in 6- month-old rats ((265 ± 20) g live weight. 30 animals were involved in the experiment, which was divided into 3 groups: control, 1st - a polylactide composite was implanted in the femur; 2nd - a new, 3-component composite material (CM) was implanted. The implants were made by 3D printing with polycomposite monofilament, which includes: polylactide - 70%, tricalcium phosphate - 20%, and hydroxylapatite - 10%

. Under general intramuscular anesthesia (aminazine 10 mg/kg and ketamine 50 mg/kg) in aseptic conditions, an incision was made of the skin and paraosseous tissues from the lateral part in the area of the metadiaphyseal part of the distal part of the femur. The bone defect (3x3 mm) was performed with a dental bur. A pin made of a composite based on polylactide was inserted into the defect. The wound was treated with a penicillin solution and sutured with Prolene-3 suture material. Additional immobilization was not performed. Glycoproteins, sialic acids, chondroitin sulfates, bilirubin, activity of acid and alkaline phosphatase, ALT, AST, GHTP, urea, and creatinine were studied in blood serum according to standard methods [4]. Statistical analysis of research data was carried out using Microsoft Excel XP and Statsoft Statistica 6.0 software packages.

Research results. Taking into account the increase in the frequency of use of implants made of composite materials in medicine in connection with the rapid development of chemistry and the improvement of production technologies, as well as with the appearance of several materials close in their properties to bone tissue, there was a need to conduct experimental studies to determine their impact on the living organism. For this purpose, several biochemical indicators reflecting the state of the metabolic processes of the animal body were investigated in the work. According to the data of the study of the blood of rats compared to the control, it was established that on the 30th day after the implantation of the composite material with PLA, there was an increase in the content of glycoproteins - by 39.9%, chondroitin sulfates - by 67.0%, and the activity of alkaline phosphatase - by 67.4 %, which has important clinical and diagnostic value for understanding the process of reparative regeneration of the femur (tabl. 1).

Table 1

Biochemical markers of blood serum of rats after implantation of composite material based on polylactide - I group (Me, 25% - 75%)

[author's development] Note: * is Wilcoxon significant compared to the control group, p<0.05;

? - probable according to Wilcoxon compared to indicators on day 30, p<0.05.

Glycoproteins play an important role in the formation of the structure of the intercellular substance, they are part of both the fibers and the amorphous substance of the connective tissue. Their spatial structure regulates the diffusion of water and low molecular weight products [5, 6]. On the 90th day (compared to the 30th), the content of glycoproteins in the blood decreased by 30.6% (reaching control), and the content of chondroitin sulfates and the activity of alkaline phosphatase increased by 12.0% and 24.9%, respectively, compared to the control. Chondroitin sulfates of connective tissue participate in the mineralization of bone tissue, contributing to the process of calcium deposition, proliferation, and exchange of chondrocytes, participate in the formation of collagen fibers, and improve blood supply in the subchondral zone of the bone [7]. The maximum intensity of metabolism is usually observed during the active period of bone structure formation.

Table 2

Biochemical markers of blood serum of rats after implantation of composite material 70% polylactide: 20% tricalcium phosphate: 10% hydroxyl apatite - II group (Me, 25% - 75%)

[author's development] Note: * is Wilcoxon significant compared to the control group, p < 0.05, ? is Wilcoxon significant compared to the 30-day values, p < 0.05.

The mineralization process of bone tissue is also associated with an increase in the activity of alkaline phosphatase, synthesized by osteoblasts, which in the damage zone increases the concentration of phosphorus ions in the structures of the extracellular matrix to the required level. Thus its content in the blood plasma is an important indicator of the process of bone reparative regeneration [8].

In the group of rats implanted with new composite material based on 70% polylactide: 20% tricalcium phosphate: 10% hydroxyl apatite compared to the control, the content of glycoproteins increased by 29.0%, chondroitin sulfates by 46.8%, and alkaline phosphatase activity by 47 .1% (tabl. 2). Thus, the content of glycoproteins in the 1st group (CM based on polylactide) in comparison with the indicators in the 2nd group (new CM) was increased by 8.4%, chondroitin sulfates and alkaline phosphatase activity - by 13.8%. On the 90th day of implantation, compared to the 30th, the content of glycoproteins decreased by 21.3%, chondroitin sulfates by 18.0%, and alkaline phosphatase activity by 15.7%. On the 90th day, compared to the 30th, only the content of chondroitin sulfates was 22.3% more.

The indicators of connective tissue exchange in the form of an increase in the concentration of glycoproteins, and chondroitin sulfates in the blood serum of rats in the study group 30 days after implantation indicate the activity of bone tissue matrix formation in the implantation area, while after 90 days the process of reparative regeneration is practically completed.

The given data show that the regenerative-inflammatory process in the bone tissue had an easier course in the second group of rats, which used a new 3- component composite material as an implant, in comparison with the use of polylactide implants.

Conclusions

The introduction of bioediting implants made of a composite based on 70% polylactide: 20% tricalcium phosphate: 10% hydroxyl apatite, in general, does not affect the somatic status of animals during all periods of the study.

The content of biochemical markers of the regenerative-inflammatory process (glycoproteins, chondroitin sulfates, alkaline phosphatase) in the blood serum of rats on the 30th day after implantation was lower precisely in the group of animals that used an implant made of a new composite material (70% polylactide: 20% tricalcium phosphate: 10% hydroxyl apatite), which is a sign of an easier course of regenerative processes of the femur.

Indicators of the functional state of the liver (ALT, AST, GHTP, bilirubin) and kidneys (creatinine, urea) in the early stages and on the 1 80th and 360th days after implantation did not change significantly in both groups of rats under study, which indicates about the absence of nephro- and hepatotoxicity of both types of bioediting implants in all periods of observation.

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