Study of pharmaceuticals affecting musculoskeletal system, in vivo and in vitro

Study of anti-inflammatory pharmacological substances:
- Study of anti-inflammatory activity in the model of chronic inflammation (adjuvant arthritis) in female Wistar rats;
- Evaluation of anti-inflammatory activity in the model of subacute inflammation (formalin-induced arthritis) in rats;
- Assessment of anti-inflammatory activity in the model of inflammation named carrageenan air pouch in rats;
- Assessment of anti-inflammatory activity in the model of inflammation – carrageenan-induced rat paw edema;
- Assessment of impact on enzymatic activity of COX-1/-2;
- Assessment of influence on enzymatic activity of 5-lipoxygenase;
- Assessment of influence on production of pro- and anti-inflammatory cytokines such as IL-1, IL-6, IL-8, FNO-?, IL-10 in cell culture and in rats;
- Assessment of influence on intracellular signaling (ss. activation of MAR-kinases: p38, JNK1/2, ERK1/2) in cell culture;
- Assessment of influence on activation of transcription factors (NF-kB, etc.) in cell culture

Assessment of PA activity in the model of experimental osteoporosis in female Wistar rats;
Osteoarthritis in rabbits:

Appearance of knee joint articular surfaces of a rabbit. Pathomorphological study

Radiograph of hind paws of a rabbit

Biochemical tests:

– CBC,

– Tumor necrosis factor alpha (TNF-?),

– Phosphorus,

– Calcium,

– Alkaline phosphatase.

Physiological tests:

– Pain sensitivity,

– Volume of an affected limb,

– Temperature of an affected limb.

Publications by topic:

1. Guseva S.I., Shikov A.N., Pozharitskaya O.N., Krishen K.L., Makarov V.G., Makarova M.N. Anti-inflammatory effect of tablets with Salvia extract using carrageenan air pouch model // Book of abstracts of the 57^thInterregional Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research. August 16-20, 2009, Geneva. Switzerland, 20097
2. Kryshen K.L., Atsapkina A.A., Akulova E.G., Rynchukovsky M.S., Makarova M.N., Pozharitskaya O.N., Shikov A.N., Makarov V.G., Dadali A. Anti-inflammatory effects of two peat distillates on the model of carrageenan air pouch in the rat. // Phytopharm. 2011. 15^thInternational congress. Book of abstracts. Nuremberg.Germany 25-27 July 2011. P.70-71
3. Makarov V.G., Makarova M.N., Kovaleva M.A., Selezneva A.I., Shikov A.N., Djachuk G.I. Evaluation of anti-inflammatory effects of peat in model of adjuvant induced artritis // Phytopharm. 2011. 15^thInternational congress. Book of Nuremberg.Germany 25-27 July 2011. P.71-72
4. Ribakova A.V., Kovaleva M.A., Kryshen K.L., Avdeeva O.I., Makarenko I.E., Makarova M.N., Makarov V.G. Antiiflamatory properties of peptide complex obtained from the cod liver (Cadidae) on the model of adjuvant arthritis in the rat// Abstracts. Phytopharm. 2012. Obzory po klinicheskoj farmacologii i lekarstvennoj terapii. -2012. –Vol. 10, № 2. –M93.
5. Rydlovskaya A.V., Makarov V.G., Makarova M.N., Tikchonov V.P. Investigation of anti- inflammatory activity of complex herbal oil extract in vitro and in vivo // Oxygen Club of California. 2006. Oxidants and antioxidants in biology. Book of abstracts. 15-18 march 2006. Santa barbara California. P.107
6. Shikov A.N., Makarova M.N., Kovaleva M.A., Pozharitskaya O.N., Makarov V.G., Djachuk G.I. Effects of peat distillates in adjuvant arthritis of rats // Abstracts 8^th Joint meeting of AFERP, ASP, GA, PSE&SIF. -Planta Med. -2012. –Vol. 78. –P. PD61.
7. Shikov A.N., Pozharitskaya O.N., Makarov V.G., Makarova M.N. Anti-inflammatory effect of Pinus sibirica oil extract in animal models //J. Nat. Med. -2008. –Vol. 62. -P. 436-440.
8. Макарова М.Н., Рыдловская А.В., Макаров В.Г. Оценка противовоспалительного действия комбинированного фитопрепарата артрофлекс и его компонентов на модели адьювантного артрита у крыс // Актуальные проблемы создания новых лекарственных препаратов природного происхождения – Фитофарм. – 2006, 27-30 июня: Матер. X международного съезда. – Санкт-Петербург, Россия. – 2006. С.279-281
9. Рыдловская А.В., Макарова М.Н., Макаров В.Г., Иванова С.А., Пожарицкая О.Н., Тихонов В.П. Оценка противовоспалительного действия комбинированного природного препарата Артрофлекс на модели каррагенинового отека у крыс линии Вистар // Вестник Санкт- Петербургской Медицинской Академии им. И.И. Мечникова.- 2006, № 3(7) С.138-141.
10. Shekunova E.V., Kashkin V.A., Muzhikyan A.A., Makarova M.N., Makarov V.G. Comparative study of two experimental models of chronic arthritis in rats // Experimental and Clinical Pharmacology. -2016. -T. 79, number 10. P. 22-29.
11. Makarova M.N., Makarov V.G. Diet-induced models of metabolic disorders. Report 7: Experimental osteoporosis // Laboratory Animals for Science. – 2019, 3. https://doi.org/10.29296/2618723X-2019-03-06 Abstract. The high incidence of osteoporosis dictates the need to find ways to effectively prevent and treat it, including in animal experiments. Of the huge number of models on different types of animals (rodents, rabbits, sheep, etc.), the most part is used by rats, which not only have a similarity of pathophysiological reactions of the skeleton with those of man, but also convenient to maintain, maintain and breed, cheap and do not have such ethical restrictions as dogs, cats and monkeys, etc. In this case, apply mainly rats wild outbred lines (Sprague Dawley, etc.). For the induction of osteoporosis, the most appropriate are complex methods that combine ovariectomy in females or orchidectomy in males, with simultaneous administration of corticosteroids against the background of a diet deficient in calcium, and sometimes in phosphorus and vitamin D. in This case, females are preferable to males due to the faster development of osteoporosis. Usually, synthetic diets containing 0.1% and less calcium are used to model osteoporosis (control animals receive a diet with 0.6–1.1% calcium). To assess the presence of osteoporosis in experimental animals should be used indicators characterizing both the intensity of bone formation (activity of alkaline phosphatase, the content of osteocalcin and procollagen I serum polypeptides), and the level of bone resorption (activity of tartrate-resistant acid phosphatase of blood serum, the content of hydroxyproline and pyridinolines in urine). In addition, it is important to the content of calcium and phosphorus in the blood serum, excretion of calcium in the urine, bone mineral density, weight of the thighs and their rupture test, as well as histomorphology of bone tissues. It is shown that the greatest changes in animals with osteoporosis are characteristic of indicators characterizing the strength of bone tissues and their weight, the activity of alkaline phosphatase, body weight, excretion of calcium in the urine, while the content of calcium and serum phosphorus does not change significantly [Full text is available in Russian].
12. Kovaleva M., Guschin Ya. Аpproval of model of chronic osteoarthrosis on rabbits . Laboratory Animals for Science. – 1991, 4. https://doi.org/10.29296/2618723X-2019-04-03 Abstract. Osteoarthrosis (OA) is a heterogeneous group of diseases of different etiology. It progresses in synovial articulation with a gradual development of the disease. Currently, osteoarthrosis (OA) is a socially significant pathology that affects the quality and life expectancy of patients. The main impairments in OA – the processes of degeneration and destruction – develop at the level of articular cartilage. In recent years, scientists’ views of the nature and mechanisms of OA development have undergone significant changes. Several decades ago, pathological changes in cartilage in OA were considered only as a consequence of natural age involution and degeneration. Currently, the key role in the development of the disease is assigned to the inflammatory process and changes in the subchondral bone. In foreign literature, the term “osteoarthritis” is often used, emphasizing the leading role of inflammation in the development of the disease. Preclinical studies make a significant contribution to the exploration of the mechanisms of OA pathogenesis. There are many methods of experimental pathology modeling, including surgical, as a result of which the changes in the tissue components of the joints are reproduced, imitating those in osteoarthrosis in humans. The purpose of this work is to test the modeling of experimental chronic osteoarthrosis using Yoshioka M. approach and to establish the possibility of using this approach to assess the pharmacological activity of drugs for the treatment of osteoarthrosis. Osteoarthrosis was induced by transection of the anterior cruciate ligament of a rabbit’s knee joint using the Yoshioka M. approach. The study was performed on 15 male rabbits, during the experiment the measurement of the circumference and pathomorphological examination of the joint was performed. Chondroitin sodium sulfate, a solution for intramuscular administration of 100 mg/ml at a dose of 6.72 mg/kg was used as a positive control drug, and Ketoprofen, capsules 320 mg at a dose of 16 mg/kg, was used as a negative control. It was shown that the transection of the anterior cruciate ligament of the rabbit’s left knee joint leads to the development of experimental pathology characterized by pronounced pathomorphological changes in the joint: the occurrence of erosions, areas of fragmentation and dissection, fibrosis of the articular capsule and ligaments. The positive control drug chondroitin sodium sulfate had a pronounced therapeutic effect, which was to reduce the circumference of the joint and it was confirmed by a pathological study. In the group of animals treated with Ketoprofen, only a decrease in the circumference of the joint was observed. Thus, the results showed that this methodological approach reproduces some aspects of the pathological changes of the joint characteristic of osteoarthrosis, and is acceptable for the assessment of the pharmacological effects of drugs intended for the pathogenetic therapy of osteoarthrosis [Full text is available in Russian].

1. Guseva S.I., Shikov A.N., Pozharitskaya O.N., Krishen K.L., Makarov V.G., Makarova M.N. Anti-inflammatory effect of tablets with Salvia extract using carrageenan air pouch model // Book of abstracts of the 57^thInterregional Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research. August 16-20, 2009, Geneva. Switzerland, 20097
2. Kryshen K.L., Atsapkina A.A., Akulova E.G., Rynchukovsky M.S., Makarova M.N., Pozharitskaya O.N., Shikov A.N., Makarov V.G., Dadali A. Anti-inflammatory effects of two peat distillates on the model of carrageenan air pouch in the rat. // Phytopharm. 2011. 15^thInternational congress. Book of abstracts. Nuremberg.Germany 25-27 July 2011. P.70-71
3. Makarov V.G., Makarova M.N., Kovaleva M.A., Selezneva A.I., Shikov A.N., Djachuk G.I. Evaluation of anti-inflammatory effects of peat in model of adjuvant induced artritis // Phytopharm. 2011. 15^thInternational congress. Book of Nuremberg.Germany 25-27 July 2011. P.71-72
4. Ribakova A.V., Kovaleva M.A., Kryshen K.L., Avdeeva O.I., Makarenko I.E., Makarova M.N., Makarov V.G. Antiiflamatory properties of peptide complex obtained from the cod liver (Cadidae) on the model of adjuvant arthritis in the rat// Abstracts. Phytopharm. 2012. Obzory po klinicheskoj farmacologii i lekarstvennoj terapii. -2012. –Vol. 10, № 2. –M93.
5. Rydlovskaya A.V., Makarov V.G., Makarova M.N., Tikchonov V.P. Investigation of anti- inflammatory activity of complex herbal oil extract in vitro and in vivo // Oxygen Club of California. 2006. Oxidants and antioxidants in biology. Book of abstracts. 15-18 march 2006. Santa barbara California. P.107
6. Shikov A.N., Makarova M.N., Kovaleva M.A., Pozharitskaya O.N., Makarov V.G., Djachuk G.I. Effects of peat distillates in adjuvant arthritis of rats // Abstracts 8^th Joint meeting of AFERP, ASP, GA, PSE&SIF. -Planta Med. -2012. –Vol. 78. –P. PD61.
7. Shikov A.N., Pozharitskaya O.N., Makarov V.G., Makarova M.N. Anti-inflammatory effect of Pinus sibirica oil extract in animal models //J. Nat. Med. -2008. –Vol. 62. -P. 436-440.
8. Макарова М.Н., Рыдловская А.В., Макаров В.Г. Оценка противовоспалительного действия комбинированного фитопрепарата артрофлекс и его компонентов на модели адьювантного артрита у крыс // Актуальные проблемы создания новых лекарственных препаратов природного происхождения – Фитофарм. – 2006, 27-30 июня: Матер. X международного съезда. – Санкт-Петербург, Россия. – 2006. С.279-281
9. Рыдловская А.В., Макарова М.Н., Макаров В.Г., Иванова С.А., Пожарицкая О.Н., Тихонов В.П. Оценка противовоспалительного действия комбинированного природного препарата Артрофлекс на модели каррагенинового отека у крыс линии Вистар // Вестник Санкт- Петербургской Медицинской Академии им. И.И. Мечникова.- 2006, № 3(7) С.138-141.
10. Shekunova E.V., Kashkin V.A., Muzhikyan A.A., Makarova M.N., Makarov V.G. Comparative study of two experimental models of chronic arthritis in rats // Experimental and Clinical Pharmacology. -2016. -T. 79, number 10. P. 22-29.
11. Makarova M.N., Makarov V.G. Diet-induced models of metabolic disorders. Report 7: Experimental osteoporosis // Laboratory Animals for Science. – 2019, 3. https://doi.org/10.29296/2618723X-2019-03-06 Abstract. The high incidence of osteoporosis dictates the need to find ways to effectively prevent and treat it, including in animal experiments. Of the huge number of models on different types of animals (rodents, rabbits, sheep, etc.), the most part is used by rats, which not only have a similarity of pathophysiological reactions of the skeleton with those of man, but also convenient to maintain, maintain and breed, cheap and do not have such ethical restrictions as dogs, cats and monkeys, etc. In this case, apply mainly rats wild outbred lines (Sprague Dawley, etc.). For the induction of osteoporosis, the most appropriate are complex methods that combine ovariectomy in females or orchidectomy in males, with simultaneous administration of corticosteroids against the background of a diet deficient in calcium, and sometimes in phosphorus and vitamin D. in This case, females are preferable to males due to the faster development of osteoporosis. Usually, synthetic diets containing 0.1% and less calcium are used to model osteoporosis (control animals receive a diet with 0.6–1.1% calcium). To assess the presence of osteoporosis in experimental animals should be used indicators characterizing both the intensity of bone formation (activity of alkaline phosphatase, the content of osteocalcin and procollagen I serum polypeptides), and the level of bone resorption (activity of tartrate-resistant acid phosphatase of blood serum, the content of hydroxyproline and pyridinolines in urine). In addition, it is important to the content of calcium and phosphorus in the blood serum, excretion of calcium in the urine, bone mineral density, weight of the thighs and their rupture test, as well as histomorphology of bone tissues. It is shown that the greatest changes in animals with osteoporosis are characteristic of indicators characterizing the strength of bone tissues and their weight, the activity of alkaline phosphatase, body weight, excretion of calcium in the urine, while the content of calcium and serum phosphorus does not change significantly [Full text is available in Russian].
12. Kovaleva M., Guschin Ya. Аpproval of model of chronic osteoarthrosis on rabbits . Laboratory Animals for Science. – 1991, 4. https://doi.org/10.29296/2618723X-2019-04-03 Abstract. Osteoarthrosis (OA) is a heterogeneous group of diseases of different etiology. It progresses in synovial articulation with a gradual development of the disease. Currently, osteoarthrosis (OA) is a socially significant pathology that affects the quality and life expectancy of patients. The main impairments in OA – the processes of degeneration and destruction – develop at the level of articular cartilage. In recent years, scientists’ views of the nature and mechanisms of OA development have undergone significant changes. Several decades ago, pathological changes in cartilage in OA were considered only as a consequence of natural age involution and degeneration. Currently, the key role in the development of the disease is assigned to the inflammatory process and changes in the subchondral bone. In foreign literature, the term “osteoarthritis” is often used, emphasizing the leading role of inflammation in the development of the disease. Preclinical studies make a significant contribution to the exploration of the mechanisms of OA pathogenesis. There are many methods of experimental pathology modeling, including surgical, as a result of which the changes in the tissue components of the joints are reproduced, imitating those in osteoarthrosis in humans. The purpose of this work is to test the modeling of experimental chronic osteoarthrosis using Yoshioka M. approach and to establish the possibility of using this approach to assess the pharmacological activity of drugs for the treatment of osteoarthrosis. Osteoarthrosis was induced by transection of the anterior cruciate ligament of a rabbit’s knee joint using the Yoshioka M. approach. The study was performed on 15 male rabbits, during the experiment the measurement of the circumference and pathomorphological examination of the joint was performed. Chondroitin sodium sulfate, a solution for intramuscular administration of 100 mg/ml at a dose of 6.72 mg/kg was used as a positive control drug, and Ketoprofen, capsules 320 mg at a dose of 16 mg/kg, was used as a negative control. It was shown that the transection of the anterior cruciate ligament of the rabbit’s left knee joint leads to the development of experimental pathology characterized by pronounced pathomorphological changes in the joint: the occurrence of erosions, areas of fragmentation and dissection, fibrosis of the articular capsule and ligaments. The positive control drug chondroitin sodium sulfate had a pronounced therapeutic effect, which was to reduce the circumference of the joint and it was confirmed by a pathological study. In the group of animals treated with Ketoprofen, only a decrease in the circumference of the joint was observed. Thus, the results showed that this methodological approach reproduces some aspects of the pathological changes of the joint characteristic of osteoarthrosis, and is acceptable for the assessment of the pharmacological effects of drugs intended for the pathogenetic therapy of osteoarthrosis [Full text is available in Russian].