Evaluation of the anesthetic efficacy of liposome encapsulated 1% ropivacaine gel for topical anesthesia in dentistry

Orientadores: Francisco Carlos Groppo, Eneida de PaulaDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: O objetivo deste estudo cruzado e cego foi avaliar a eficácia da anestesia tópica de ropivacaína a 1% (forma lipossomada ou não) em Odontologia. Trinta voluntários foram submetidos à anestesia tópica com quatro anestésicos, em oito sessões (tratamentos) determinadas aleatoriamente: 20mg de gel de ropivacaína a 1% -Ropi-20, 60mg de gel de ropivacaína a 1% -Ropi-60, 20mg de gel de ropivacaína a 1% encapsulada em lipossomas -RopiLipo-20, 60mg de gel de ropivacaína a 1% encapsulada em lipossomas -RopiLipo-60, 20mg da mistura eutética de lidocaína a 2.5% e prilocaína a 2.5% (EMLA cream AstraZeneca®) - EMLA-20, 60mg de EMLA cream -EMLA-60, 20mg de gel de benzocaína a 20% (Benzotop® DFL) -Benzo-20 e 60mg de gel de benzocaína a 20% -Benzo-60. Cada tratamento foi aplicado no fundo de sulco da região de canino superior direito durante dois minutos; o intervalo entre os tratamentos foi de uma semana. Foram avaliadas a anestesia pulpar, por meio de estímulo elétrico (pulp tester); a dor durante a punção por meio das escalas analógica visual (EAV) e de 11 pontos em caixa (E11); e a anestesia em tecido mole por meio de estímulo físico. A influência do anestésico na resposta pulpar foi avaliada durante 20 minutos após a aplicação do anestésico tópico. Não houve diferença estatisticamente significante entre os tratamentos com relação às escalas EAV e E11 (p>0.05). A duração da anestesia em tecido mole com o EMLA-60 e RopiLipo-60 foi maior (p0.05). Nenhum dos tratamentos avaliados exerceu efeito anestésico sobre o tecido pulpar. Assim, a ropivacaína a 1% encapsulada ou não em lipossomas apresentaram eficácia semelhante em reduzir a dor à punção em comparação aos outros anestésicos avaliados, no entanto, nas condições avaliadas, nenhum dos anestésicos exerceu influência sobre a resposta pulparAbstract: The objective of this blind and crossover study was to evaluate the efficacy of 1% ropivacaine (liposomal and nonliposomal) for topical anesthesia in Dentistry. Thirty healthy volunteers randomly received the following treatments in eight sessions: 20mg of 1% ropivacaine -Ropi-20, 60mg of 1% ropivacaine -Ropi-60, 20mg of liposome encapsulated 1% ropivacaine -RopiLipo-20, 60mg of liposome encapsulated 1% ropivacaine -RopiLipo-60, 20mg of the eutectic mixture of 2.5% lidocaine and 2.5% prilocaine (EMLA cream AstraZeneca®) -EMLA-20, 60mg of EMLA cream -EMLA-60, 20mg of 20% benzocaine gel (Benzotop® DFL) -Benzo-20 and 60mg of 20% benzocaine gel -Benzo-60. At different sessions each treatment was applied in the maxillary-buccal fold of the right canine for two minutes; one week of interval between treatments. Pulpal anesthesia was evaluated using an electrical pulp tester; pain during needle insertion by using a visual analogue scale (VAS) and 11-point box scale (BS-11) and soft tissue anesthesia by using a pinprick test. The influence on pulpal response was assessed for 20 minutes after topical application. There were no statistically significant differences among treatments considering VAS and BS-11 scores (p=0.177 and p=0.179, respectively). RopiLipo-60 and EMLA-60 provided a statistically significant longer duration of topical anesthesia than the other topical agents (p<0.05), however, RopiLipo-60 was not significantly different from EMLA-60 (p>0.05). There was no influence on the pulpal response provided by the topical anesthetics evaluated. Liposome and nonliposomal ropivacaine gel at 1% concentration showed similar efficacy for topical anesthesia in oral mucosa when compared to the other topical anesthetics. However, none of the topical anesthetics evaluated were effective in inducing pulpal anesthesia under the evaluated conditionsMestradoFarmacologia, Anestesiologia e TerapeuticaMestre em Odontologi

Anesthetic efficacy and plasmatic concentration of liposome-encapsulated ropivacaine in dental anesthesia

Orientadores: Maria Cristina Volpato, Eneida de Paula, Francisco Carlos GroppoTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: Uma nova forma farmacêutica de anestésico local, encapsulado em lipossomas, vem sendo estudada na Medicina e mais atualmente em Odontologia. Os objetivos deste trabalho foram avaliar a eficácia anestésica em anestesia tópica e infiltrativa e os parâmetros farmacocinéticos da ropivacaína encapsulada em lipossomas, em 4 estudos, cruzados, duplo-cegos e com ordem de aplicação aleatória, com intervalo de 1 semana entre as aplicações. Capítulo 1: foram comparadas a eficácia da anestesia tópica e a influência na resposta pulpar da ropivacaína 2% encapsulada em lipossomas (RL2), Benzocaína 20% ( - B20), gel placebo lipossomal (PL) e gel placebo (P) aplicados em mucosa vestibular dos incisivos laterais superiores, em 40 voluntários. RL2 foi tão eficaz quanto B20 em reduzir dor à punção e na duração de anestesia em tecidos moles (p>0,05) e ambas foram superiores às formulações PL e P (p<0,05). Nenhuma das formulações exerceu influência na resposta pulpar. Capítulo 2: ropivacaína 2% encapsulada em lipossomas (RL2), ropivacaína 1% encapsulada em lipossomas (RL1), creme de lidocaína 2,5% e prilocaína 2,5% (EMLA) e gel placebo lipossomal (PL) foram avaliados quanto à eficácia em reduzir dor à punção e à injeção de anestésico local, quando aplicados topicamente na região palatina do canino superior esquerdo. O EMLA foi mais efetivo em diminuir a dor à punção (p0,05). Nenhuma das formulações lipossomais foi eficaz como anestésico tópico na mucosa palatina. Capítulo 3: foram injetados, no fundo de sulco vestibular do canino superior direito, 1,8mL de ropivacaína 0,5% encapsulada em lipossomas (RLipo), ropivacaína 0,5% com epinefrina 1:200.000 (Repi), ropivacaína a 0,5% (R) e lidocaína 2% com epinefrina 1:100.000 (Lepi), em 40 voluntários. Foram avaliadas latência e duração da anestesia pulpar por aplicação de estímulo elétrico e em tecidos moles por estímulo de pressão. Não houve diferença estatística entre os anestésicos com relação ao tempo de latência. Repi e Lepi apresentaram maior tempo de anestesia pulpar quando comparados à RLipo e R (p0,05) entre os parâmetros farmacocinéticos avaliados entre as duas soluções anestésicas. Conclusão geral: Não há vantagem no uso da ropivacaína 0,5% encapsulada em lipossomas em técnica infiltrativa ou 1 e 2% em anestesia tópica em mucosa palatina. Em mucosa vestibular, por apresentar eficácia semelhante à da benzocaína 20%, a ropivacaína 2% encapsulada em lipossomas pode ser uma opção a esse anestésico. A ropivacaína encapsulada em lipossomas apresenta perfil farmacocinético semelhante ao da ropivacaína com epinefrina.Abstract: A new pharmaceutical formulation of local anesthetic, liposome encapsulated, has been studied in medicine and recently in dentistry. The aims of the present study were to evaluate anesthetic efficacy in topical and infiltration anesthesia, and pharmacokinetic parameters of liposome-encapsulated ropivacaine in 4 random, crossed and double-blind studies, with a one week interval between sections. Chapter 1: liposome-encapsulated 2% ropivacaine (RL2), 20% Benzocaine (B20), liposomal placebo (PL) and placebo (P) were compared in relation to the efficacy of topical anesthesia and influence on pulpal response after topical application in the buccal fold of the upper lateral incisors, in 40 volunteers. RL2 was as efficacious as B20 in reducing pain during needle insertion and concerning soft tissue anesthesia (p>0.05) and both agents were better than PL e P formulations (p0.05). None of the formulations was effective as a topical anesthetic in the palatine mucosa. Chapter 3: forty volunteers received 1.8mL of liposomeencapsulated 0.5% ropivacaine (RLipo), 0.5 % ropivacaine with 1:200,000 epinephrine (Repi), 0.5% ropivacaine (R) and 2% lidocaine with 1:100,000 epinephrine (Lepi), as an infiltration injection in the buccal fold of the right maxillary canine region. The onset and duration of pulpal anesthesia were evaluated through electric stimuli application and in soft tissue by pressure stimuli. No difference in onset of anesthesia was observed among anesthetic formulations (p>0.05). Repi and Lepi showed longer pulpal anesthesia when compared to RLipo and R (p0.05) among pharmacokinetics parameters between the two anesthetic formulations. Final conclusion: There is no advantage in the use of liposome-encapsulated 0.5% ropivacaine in infiltration anesthesia or liposome-encapsulated 1 and 2% ropivacaine in topical anesthesia in palatal mucosa. In the buccal mucosa, as it showed similar efficacy of 20% benzocaine, liposome-encapsulated 2% ropivacaine can be an option to this anesthetic. Liposome-encapsulated ropivacaine and ropivacaine with epinephrine showed similar pharmacokinetic.DoutoradoFarmacologia, Anestesiologia e TerapeuticaDoutor em Odontologi

Pharmacokinetic profile of liposome-encapsulated ropivacaine after maxillary infiltration anaesthesia

The aim of this study was to determine the pharmacokinetic parameters of liposomal ropivacaine after dental anesthesia in 14 healthy volunteers. In this randomized, double-blind and crossover study, the volunteers received maxillary infiltration of liposome-encapsulated 0.5% ropivacaine and, 0.5% ropivacaine with 1:200,000 epinephrine in two different sessions. Blood samples were collected before and after (from 15 to 1440 min) the administration of either ropivacaine formulation. HPLC with UV detection was used to quantify plasma ropivacaine concentrations. The pharmacokinetic parameters AUC(0-24) (area under the plasma concentration x time curve from baseline to 24 h), AUC(0-infinity) (area under the plasma concentration-time curve from baseline to infinity), C-max (maximum drug concentration), CL (renal clearance), T-max (maximum drug concentration time), t(1/2) (elimination half-life) and Vd (volume of distribution) were analyzed using the Wilcoxon signed-rank test. No differences (p > 0.05) were observed between both formulations for any of the pharmacokinetic parameters evaluated and plasma ropivacaine concentrations, considering each period of time. Both formulations showed similar pharmacokinetic profiles, indicating that the liposomal formulation could be a safer option for use of this local anesthetic, due to the absence of a vasoconstrictor

Máscaras caseiras na pandemia de COVID-19: recomendações, características físicas, desinfecção e eficácia de uso

Objective: To describe the recommendations, physical characteristics, methods of disinfection and the effectiveness of using homemade masks in reducing the transmission of COVID-19. Methods: Search was performed in the MEDLINE, SciELO and Google Scholar databases, in addition to the official recommendations for use. Results: 31 references were included. The filtration capacity of tissues varied between 5 and 98%. 100% cotton fabrics in two or three layers showed filtration efficiency between 70 and 99%, in vitro. Homemade masks, surgical and respirators showed breathability between 2.2 to 3.0 Pascal. The ability to reduce the spread of microorganisms by people wearing home masks was three times less than when using surgical masks, but it was superior to not wearing masks. Conclusion: The breathability of homemade masks may be adequate, while the filtration efficiency seems to be inferior to surgical masks, but superior to no mask use. There is no evidence to support the efficacy and effectiveness of homemade masks.Objetivo: Descrever as recomendações, características físicas, métodos de desinfecção e eficácia de uso de máscaras caseiras na redução da transmissão da COVID-19. Métodos: Realizou-se busca nas bases de dados MEDLINE, SciELO e Google Scholar, além das recomendações oficiais de uso. Resultados: Foram incluídas 31 referências. A capacidade de filtração de tecidos variou entre 5 e 98%. Tecidos 100% algodão em duas ou três camadas apresentaram eficácia de filtração entre 70 e 99% em estudos in vitro. Máscaras caseiras, cirúrgicas e respiradores apresentaram respirabilidade entre 2,2 e 3,0 Pascal. A capacidade de redução da propagação de microrganismos por pessoas usando máscaras caseiras foi três vezes menor do que ao usar máscaras cirúrgicas, embora tenha sido superior ao não uso de máscaras. Conclusão: A respirabilidade de máscaras caseiras mostrou-se adequada, enquanto a eficácia de filtração parece ser inferior à das máscaras cirúrgicas, mas superior a não usar máscara. Não há evidências que respaldem a eficácia e efetividade das máscaras caseiras.&nbsp

Pharmacokinetic Profile of Liposome-Encapsulated Ropivacaine after Maxillary Infiltration Anaesthesia

O objetivo do presente estudo foi determinar os parâmetros farmacocinéticos da ropivacaína encapsulada em lipossomas após anestesia local em 14 voluntários sadios. Neste estudo randomizado, cruzado e duplo-cego, os voluntários receberam anestesia infiltrativa na maxila de ropivacaína a 0,5% encapsulada em lipossomas e ropivacaína 0,5% com epinefrina a 1:200.000 em duas sessões distintas. Amostras de sangue foram coletadas antes e após (de 15 a 1440 min) a administração das formulações de ropivacaína. A quantificação da concentração plasmática de ropivacaína foi feita por meio de HPLC com detecção por UV. Os parâmetros farmacocinéticos AUC 0-24 (área sob a curva de concentração × tempo do tempo 0 até 24 horas) , AUC 0-∞ (área sob a curva de concentração x tempo do tempo 0 até o infinito), C max (concentração máxima da droga), CL (clearance renal), T max (tempo em que ocorre a concentração máxima); t 1/2 (meia vida de eliminação) e V d (volume de distribuição) foram analisados pelo teste de Wilcoxon. Nenhuma diferença (p &gt; 0,05) foi observada entre as duas formulações em cada parâmetro farmacocinético avaliado e as concentrações plasmáticas de ropivacaína, considerando cada período de tempo. Ambas as formulações apresentaram perfil farmacocinético semelhante, indicando que a formulação lipossomal poderia ser uma opção mais segura para o uso deste anestésico local, devido à ausência de vasoconstritor. The aim of this study was to determine the pharmacokinetic parameters of liposomal ropivacaine after dental anesthesia in 14 healthy volunteers. In this randomized, double-blind and crossover study, the volunteers received maxillary infiltration of liposome-encapsulated 0.5% ropivacaine and, 0.5% ropivacaine with 1:200,000 epinephrine in two different sessions. Blood samples were collected before and after (from 15 to 1440 min) the administration of either ropivacaine formulation. HPLC with UV detection was used to quantify plasma ropivacaine concentrations. The pharmacokinetic parameters AUC 0-24 (area under the plasma concentration × time curve from baseline to 24 h), AUC 0-∞ (area under the plasma concentration-time curve from baseline to infinity), C max (maximum drug concentration), CL (renal clearance), T max (maximum drug concentration time), t 1/2 (elimination half-life) and Vd (volume of distribution) were analyzed using the Wilcoxon signed-rank test. No differences (p &gt; 0.05) were observed between both formulations for any of the pharmacokinetic parameters evaluated and plasma ropivacaine concentrations, considering each period of time. Both formulations showed similar pharmacokinetic profiles, indicating that the liposomal formulation could be a safer option for use of this local anesthetic, due to the absence of a vasoconstrictor. Keywords: ropivacaine, local anesthesia, dentistry, liposome, pharmacokinetic Pharmacokinetic Profile of Liposome-encapsulated Ropivacaine after Maxillary Infiltration Anaesthesia J. Braz. Chem. Soc. 1946 Introduction Prolonged-action local anesthetic is required when postoperative pain and discomfort are expected, especially after major surgical procedures. 1 In many countries, bupivacaine, the racemic mixture of S-and D-bupivacaine, is the only long-acting local anesthetic available in dental practice. Ropivacaine (RVC), another long-acting local anesthetic of the cyclic aminoamide family, is synthesized in the S-enantiomer form and presents lower toxicity to the cardiovascular and central nervous systems, compared to bupivacaine. 2 Traditionally, most of local anesthetic formulations are administered together with a vasoconstrictor in order to increase anesthesia duration and to reduce systemic absorption rate. 3 It was recently demonstrated that the use of these formulations increased, especially those containing epinephrine 1:100.000. 5 Alternative drug delivery systems, such as liposomes, have been used to prolong the duration of action of many drugs, including local anesthetics. 6-10 These vesicles are nontoxic and nonimmunogenic because their components (phosphatidyl choline and cholesterol) are also found in biological membranes. 17 Previous authors have shown that liposomal encapsulation of bupivacaine altered its pharmacokinetic profile after extradural injection in rabbits, resulting in lower concentrations of the drug in plasma, liver and myocardium. 18 Grant and co-workers 16 observed that when encapsulated in liposomes, bupivacaine remained at the injection site for a significantly longer period of time after subcutaneous injection in mice. Attempting to simulate an accidental intravascular injection of a local anesthetic, Boogaerts and coworkers 8 assessed the acute CNS (central nervous system) and cardiac toxicities induced by intravenous infusion in rabbits of 0.25% bupivacaine, with and without epinephrine (1:200,000), compared to liposomeencapsulated bupivacaine. They demonstrated a reduction of CNS and cardiac toxicities using liposome-encapsulated bupivacaine. The addition of epinephrine to the plain solution did not decrease the CNS and cardiac toxicities induced by bupivacaine. It was recently demonstrated in animal studies, which used sciatic and infraorbital nerve blockades, that encapsulation of ropivacaine into unilamellar vesicles increased the duration and the intensity of analgesic effects. 6 Although long-acting local anesthetics are normally used in low doses in dentistry, high doses may be required for removal of four impacted third molars in a single session. The present study is the first attempt to measure the pharmacokinetic parameters of ropivacaine after maxillary infiltration anesthesia using a liposome-encapsulated formulation in healthy volunteers. The pharmacokinetic parameters of an RVC formulation containing epinephrine (vasoconstrictor) were also assessed for comparison. Experimental Materials RVC hydrochloride was donated by Cristalia Prod. Quim. Farm. Ltda (Itapira, SP, Brazil). Egg phosphatidylcholine (EPC), cholesterol (Ch) and α-tocopherol (α-T) were purchased from Sigma Chemical Company (St Louis, MO). All other reagents used were of analytical grade. RVC-liposome formulation The liposomal RVC formulations were prepared as described by Araújo and co-workers. 10 The following liposomal characterization was previously determined by Araújo and co-workers. Subjects This research was approved by the Ethical Committee of Piracicaba Dental School, University of Campinas (approval #164/2006). Fourteen healthy volunteers (seven males and seven females) aged 24 (± 3.1) years old were selected, and signed a written informed consent. The volunteers presented no systemic or oral disorders, had no history of allergy to any of the local anesthetics used, and were not taking any medication, as determined by oral questioning and by their documented health histories. Prior to the beginning and right after the end of the study, all the subjects were submitted to laboratory tests to confirm that they were in good health and that the females were not pregnant. The same tests were performed at the end of the study to ensure that all results previously obtained were not altered by the anesthetic formulations tested. These tests included cross-reactive protein, blood-hemoglobin, lymphocyte count, platelet count, erythrocyte sedimentation rate, serum (S)-sodium, S-potassium, S-chloride, S-albumin, S-alkaline phosphate, S-gamma-glutamyl-transferase, S-aspartate transaminase, S-alanine transaminase, S-creatine, plasma glucose, urea, cholinesterase, total protein, bilirubin, uric acid, urine glucose, urine leukocyte count, urine protein, and urine hemoglobin. Serology tests for human immunodeficiency virus and hepatitis B and C were also performed. Female subjects underwent a urine bHCG pregnancy test. Ambulatory procedures Anesthetic procedures In this double-blind and crossover study, the volunteers randomly received 1.8 mL of 0.5% ropivacaine with 1:200,000 epinephrine, and liposome-encapsulated 0.5% ropivacaine, for infiltration anesthesia at the apex of the right maxillary canine, in two different sessions spaced one week apart. Ropivacaine with 1:200,000 epinephrine was obtained by simple dilution of the commercially available solution of ropivacaine (Naropin ® 10 mg mL -1 , AstraZeneca, São Paulo, Brazil) immediately before application. Under sterile conditions, 5 mL of 1% ropivacaine was diluted with 5 mL of 1:100,000 (v/v) epinephrine (Drenalin ® , Ariston Ind. Quim. Farm. Ltda, São Paulo, SP, Brazil). The local anesthetics (1.8 mL) were placed into coded sterile 3 mL Luer-Lok syringes (Becton Dickinson, Curitiba, Brazil) fitted with disposable needles (30G, one-inch, Becton-Dickinson Company, Franklin Lakes, NJ, USA). After topical anesthesia on the injection site with 20% benzocaine, the formulations were injected at the maxillary buccal fold of the right-canine region at an injection rate of 1 mL min -1 . The same operator performed the maxillary infiltration anesthesia in all the subjects. Blood sampling and drug analysis Blood samples (4.5 mL) from a forearm vein were collected with a heparinized cannula before and 15, Pharmacokinetic and statistical analyses The following pharmacokinetic parameters were evaluated by computer software (PK Solutions, noncompartmental pharmacokinetics data analysis, 2001; Summit Research Services, Montrose, CO, USA): C max (maximum drug concentration); T max (maximum drug concentration time); AUC 0-24 , (area under the plasma concentration-time curve from baseline to 24 h); AUC 0-∞ (area under the plasma concentration-time curve from baseline to infinity); CL (renal clearance); t 1/2 (elimination half-life) and Vd (volume of distribution). Statistical analysis was performed using the Student&apos;s t-test in order to compare the ropivacaine concentrations between the groups at each time interval. Pharmacokinetic parameters were compared using the Wilcoxon signed-rank test. The significance level was set at 5%, and the tests were performed with BioEstat 5.0 (Fundação Mamirauá, Belém, PA, Brazil) Results and Discussion In an earlier study, Araújo and co-workers 6 demonstrated that the size distribution of liposomal formulations containing RVC presented two modes, one with a maximum at 371 nm (85%), and another with a peak at 128 nm (15%). The efficiency of encapsulation was around 24%, which was sufficient to modify the release profile of the pharmaceutical, with a reduction of the release rate over a one-hour period from 76 to 58%. In the same study it was also shown that, compared to RVC alone, the liposomal RVC formulation increased the duration and intensity of analgesic effects in sciatic and infraorbital nerve blocking experiments. Extending the earlier work of Araújo and co-workers 6 here we report on the first attempt to assess the pharmacokinetic parameters of ropivacaine after maxillary infiltration anesthesia using a liposome-encapsulated ropivacaine formulation in healthy volunteers, comparing the results with a commercial RVC formulation containing epinephrine vasoconstrictor. The calibration curve for determination of plasma ropivacaine ( 22 The detection limit for ropivacaine observed in our study (30 ng mL -1 ) was close to the limit observed by those authors (25 ng mL -1 )

Meningkatkan Hasil Belajar Siswa Kelas IV MIS Tompo Melalui Pemanfaatan Lingkungan Sekolah Sebagai Sumber Belajar IPA

Penelitian ini bertujuan untuk meningkatkan hasil belajar siswa pada mata IPA di Kelas IV MIS Tompo melalui Pemanfaatan Lingkungan Sekolah sebagai Sumber Belajar. Penelitian ini merupakan penelitian tindakan kelas (PTK), yang dilaksanakan sebanyak dua siklus, setiap siklus terdiri dari perencanaan, pelaksanaan tindakan, observasi dan refleksi yang mengacu pada model KemmisMc Taggart. Data yang diambil adalah data kualitatif dan kuantitatif. Data kualitatif diperoleh dari hasil aktifitas siswa dan guru, sedangkan data kuantitatif diperoleh dari hasil evaluasi disetiap siklus. Hasil siklus I menunjukkan bahwa aktifitas guru adalah: 82,85% dikategorikan baik sedang aktifitas siswa adalah, 71, 1% dikategorikan cukup, dengan ketuntasan belajar Klasikal 62,5%,. Hasil siklus II menunjukkan bahwa aktifitas guru 92,85%) dengan kategori sangat baik, sedang aktifitas siswa 82,2 % dikategorikan baik, dengan persentase daya serap klasikal sebesar 86,3%. Dengan demikian maka dapat dikatakan bahwa pemanfaatan lingkungan sekolah sebagai sumber belajar dapat meningkatkan hasil belajar IPA siswa Kelas IV MIS Tompo Kec. Taopa

Figure S4

Filled board of Stage 4 of the 4-stage educational puzzle game, developed for teaching the physiology of glomerular filtration.</p

Figure S2

Filled board of Stage 2 of the 4-stage educational puzzle game, developed for teaching the physiology of glomerular filtration.</p

Figure S3

Filled board of Stage 3 of the 4-stage educational puzzle game, developed for teaching the physiology of glomerular filtration.</p